Source: UNIVERSITY OF NEBRASKA submitted to NRP
NUTRIENT BIOAVAILABILITY--PHYTONUTRIENTS AND BEYOND
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0216251
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
W-2002
Project Start Date
Oct 1, 2008
Project End Date
Sep 30, 2013
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF NEBRASKA
(N/A)
LINCOLN,NE 68583
Performing Department
Nutritional & Health Sciences
Non Technical Summary
PURPOSE: The long-term objective of the Nebraska investigator is to elucidate mechanisms by which biotinylation of histones participates in gene regulation and genome stability. PROCEDURES: CONTRIBUTION 1: To generate, test, and validate new antibodies to novel histone biotinylation sites H2AK9bio, H2AK13bio, H3K4bio, H3K9bio, H3K18bio, and H4K8bio. CONTRIBUTION 2: To characterize the roles of H2AK9bio, H2AK13bio, H3K4bio, H3K9bio, H3K18bio, and H4K8bio in gene regulation, heterochromatin structures, and repeat regions in mammalian chromatin. CONTRIBUTION 3: To identify effects of biotin nutritional status on histone biotinylation and, therefore, gene regulation and genome stability. CONTRIBUTION 4: To collaborate with investigators from this Multi-State Project to identify interactions between chromatin methylation events and histone biotinylation, and to identify bioactive food compounds that affect the activity of holocarboxylase synthetase. OUTCOMES: (i) At least two peer-reviewed publications will result directly from this research. In addition, at least two grant proposals will be submitted to federal funding agencies based on the proposed work. (ii) At least two graduate students will be trained in molecular biology techniques during the course of the proposed studies. Two graduate students (Gaganpreet Kaur Mall and Valerie Pestinger) have already been recruited. (iii) The development of holocarboxylase synthetase inhibitors might lead to patent submissions. PROJECTED IMPACTS: (i) The proposed research will contribute to the workforce development in the State of Nebraska by recruiting and training highly qualified students and laboratory staff, and will offer mentoring opportunities for postdoctoral fellows. (ii) The proposed research will enhance UNL's recognition by peers, due to publication of this research in top-tier journals. (iii) Previous research in this laboratory has generated patent applications and has resulted in license agreements with industry. Hence, the proposed research is likely to promote the economic development in the State of Nebraska. (iv) The proposed research is likely to generate external funding for the university. This will result in increased recruitment of graduate students. (v) Graduate classes taught by Dr. Zempleni (Molecular Nutrition and Molecular Nutrition Techniques) will be updated based on discoveries made in the research proposed here. (vi) The proposed research will yield novel insights into human biotin requirements, and decreasing cancer risk by optimal nutrition. We will present our findings to dietitians and the lay public (talks, newspaper articles). We anticipate that this will result in improved biotin nutrition.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70250101010100%
Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components;

Subject Of Investigation
5010 - Food;

Field Of Science
1010 - Nutrition and metabolism;
Goals / Objectives
Determine the bioavailability (absorption, distribution, metabolism, elimination) of nutrients and other food components and their environmental and genetic determinants Evaluate the bioactivity of nutrients and other food components in order to elucidate their underlying protective mechanisms.
Project Methods
CONTRIBUTION 1: Polyclonal antibodies against H2AK9bio, H2AK13bio, H3K4bio, H3K9bio, H3K18bio, and H4K8bio will be raised as described for our previous studies. Antibody titers and specificities in pre- and post-immune sera will be tested by using synthetic peptides "targets" and "non-targets"), bulk extracts of human histones, biotin-depleted histones, and competition studies between histones and synthetic peptides. All these methods are routinely used in our laboratory. CONTRIBUTION 2: The relative enrichment of biotinylated histones in heterochromatin (repeats regions), repressed chromatin, de-repressed chromatin, and transposable elements will be quantified by chromatin immunoprecipitation assay and real-time PCR. Both wild-type and holocarboxylase synthetase knockdown (negative control) cells will be used. Relative enrichment data will be interpreted in combination with gene expression data obtained by reverse transcriptase real-time PCR. We have extensive experience with these methods and have already generated the holocarboxylase synthetase knockdown cells. CONTRIBUTION 3: Human cells will be cultured in biotin-defined media, representing plasma biotin concentrations as observed in biotin-deficient individuals, normal individuals, and users of biotin supplements. Effects of biotin supply on histone biotinylation in distinct loci of the human genome will be quantified by chromatin immunoprecipitation assay and real-time PCR. We will use the cytogenetics facility at the University of Nebraska Medical Center to investigate whether decreased histone biotinylation in biotin-deficient cells increases the frequency of chromosomal abnormalities. These analytical procedures are routinely used in our laboratory. CONTRIBUTION 4: In previous studies we have demonstrated that the relative enrichment of H4K12bio at transposable elements depends on DNA methylation in human cells. In these previous studies, DNA methylation marks were erased by treatment with azacytidine, and the relative enrichment of H4K12bio at transposable elements was quantified by chromatin immunoprecipitation assay. Here, we will use an analogous approach to study effects of azacytidine treatment on the relative enrichments of H2AK9bio, H2AK13bio, H3K4bio, H3K9bio, H3K18bio, and H4K8bio. We also intend to make use of a transgenic mouse from the University of California PI, in which DNA methylation is substantially decreased. In addition we will seek to identify bioactive food compounds that affect the activity of holocarboxylase synthetase, which is the enzyme that mediates histone biotinylation. We have already produced bioactive recombinant holocarboxylase synthetase and developed an assay system for this enzyme; some candidate compounds that might affect holocarboxylase synthetase activity have been tentatively identified.

Progress 10/01/08 to 09/30/13

Outputs
Target Audience: My target audience includes scientists, federal officials from funding agencies, graduate students, and the lay public. I have reached my target audience using the following mechanisms: (1) publications in peer-reviewed journals; (2) presentations at local, national, and international conferences; (3) website. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Four postdoctoral fellows, seven graduate students, and 5 undergraduate students have been trained. How have the results been disseminated to communities of interest? Eleven peer-reviewed papers, four book chapters, one book, one review article, and nine symposia proceedings have been published and submitted in the final year of this project. I hosted the 2013 W2002 multistate group meeting at which members of the group shared their research discoveries. Biosketches and research interests and findings of multistate group investigators are displayed on the nutrigenomics.unl.edu website in Nebraska. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We have created numerous antibodies against biotinylated histones. We have shown that the roles of biotin in gene repression are due to physical interactions between holocarboxylase synthetase (HLCS) and other chromatin proteins and that histone biotinylation sites are mere marks for HLCS docking sites in human chromatin. We demonstrated that biotin, methyl donors, HLCS, and chromatin proteins synergize in the repression of repeats in mammalian chromatin, thereby contributing toward genome stability and low cancer risk. This project included collaborations with more than 10 scientists that can be linked to publications, in addition to numerous other collaborations.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Eng WK, Giraud D, Schlegel VL, Wang D, Lee BH, Zempleni J. Identification and assessment of markers of biotin status in healthy adults. Br J Nutr 110:321-329, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Singh MP, Wijeratne SSK, Zempleni J. Biotinylation of lysine 16 in histone H4 contributes toward nucleosome condensation. Arch Biochem Biophys 529:105-111, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Camara Teixeira D, Malkaram SA, Zempleni J. Enrichment of meiotic recombination hotspot sequences by avidin capture technology. Gene 516:101-106, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xue J, Wijeratne SSK, Zempleni J. Holocarboxylase synthetase synergizes with methyl CpG binding protein 2 and DNA methyl transferase 1 in the transcriptional repression of long terminal repeats. Epigenetics 8:504-511, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Li Y, Hassan YI, Moriyama H, Zempleni J. Holocarboxylase synthetase interacts physically with euchromatic histone-lysine N-methyltransferase, linking histone biotinylation with methylation events. J Nutr Biochem 24:1446-1452, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xia M, Malkaram SA, Zempleni J. Three promoters regulate the transcriptional activity of the human holocarboxylase synthetase gene. J. Nutr Biochem 24: 1963-1969, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xue J, Zempleni J. Epigenetic synergies among biotin, folate, and holocarboxylase synthetase in the regulation of pro-inflammatory cytokines and repeats. Scand J Immunol 78: 419-425, 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xue J, Zhou J, Zempleni J. Holocaboxylase synthetase catalyzes biotinylation of heat shock protein 72, thereby inducing RANTES expression in HEK293 cells. Am J Physiol Cell Physiol 305:C1240-C1245, 2013
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Baier SR, Zbasnik R, Schlegel V, Zempleni J. Off target effects of sulforaphane include the de-repression of long-terminal repeats through histone acetylation . J Nutr Biochem 25:665-668, 2014
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Liu D, Zempleni J. Holocarboxylase synthetase interacts physically with the nuclear receptor corepressor, histone deacetylase 1, and a novel splicing variant of histone deacteylase 1 to repress repeats. Biochem J 461:477-486, 2014
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Li Y, Malkaram SA, Zhou J, Zempleni J. Lysine biotinylation and methionine oxidation in the heat shock protein HSP60 synergize in the elimination of reactive oxygen species in human cell cultures. J Nutr Biochem (in press)
  • Type: Book Chapters Status: Published Year Published: 2012 Citation: Zempleni J, Wijeratne SSK, Kuroishi T. Chapter 23: Biotin. In: Present Knowledge in Nutrition. Erdman JW Jr (ed.), 10th edition. International Life Sciences Institute, Washington, DC, 2012, pp. 587-609 (http://lto.libredigital.com/?9780470963104-Erdman_199s4l)
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Zempleni J, Liu D, Xue J. Nutrition, histone epigenetic marks, and disease. In: Epigenomics in Health and Disease. Jirtle, RL, Tyson F (eds.), Springer, Heidelberg, Germany, 2013, pp. 197-217
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Zempleni J, Liu D, Camara Teixeira D, Singh MP. Mechanisms of gene transcriptional regulation through biotin and biotin-binding proteins in mammals. In: Vitamin-binding Proteins  Their Functional Consequences. Dakshinamurti D, Dakshinamurti S (eds.), Taylor & Francis, Boca Raton, FL, 2013 pp. 219-228
  • Type: Books Status: Published Year Published: 2013 Citation: Zempleni J, Cordonier EL, Baier SR, Xue J. Vitamins, Bioactive Food Compounds, and Histone Modifications. In: Handbook of Vitamins. Zempleni J, Suttie JW, Gregory JF III, Stover PJ (eds), 5th edition. Taylor and Francis, Inc., Boca Raton, FL, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Camara D, Malkaram SA, Zempleni J. Enrichment of meiotic recombination hotspot sequences by avidin capture technology. Experimental Biology Meeting; Boston, MA, April 23, 2013
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2014 Citation: Zempleni J, Barshop B, Cordonier EL, Baier SR. Disorders of biotin metabolism. In: The Online Metabolic & Molecular Bases of Inherited Disease. Editors: Valle, Baudet, Vogelstein, Kinzler, Antonarakis, Ballabio, Scriver (Emeritus), Childs (Emeritus), Sly (Emeritus), Bunz (Parts Editor), Gibson (Parts Editor), Mitchell (Parts Editor). McGraw Hill.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Zempleni J, Liu D. Teixeira Camara D, Cordonier EL. Novel roles of holocarboxylase synthetase in gene regulation and intermediary metabolism. Nutr Rev 72:369-376, 2014
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Baier SR, Schlegel VL, Zempleni J. Off Target Effects of Dietary Sulforaphane. College of Education and Human Sciences Research Fair; Lincoln, NE, November 2, 2012
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Xue J, Zempleni J. Epigenetic synergies between methyl donors and biotin in gene repression are mediated by holocarboxylase synthetase (HLCS). Experimental Biology Meeting; Boston, MA, April 23, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Baier SR. Zbasnik R, Schlegel VL, Zempleni J. Dietary sulforaphane elicits off-target effects at loci coding for long terminal repeats in lymphocytes from healthy adults and in IMR-90 fibroblast cultures, possibly impairing genome stability. Experimental Biology Meeting; Boston, MA, April 23, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhou J, Wijeratne SSK, Zempleni J. Biotinylation of the c-myc promoter binding protein MBP-1 decreases c-myc expression in mammary carcinoma MCF-7 cells. Experimental Biology Meeting; Boston, MA, April 21, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cordonier EL, Teixeira Camara D, Han Z, Pannier AK, Zempleni J. Acetyl-CoA carboxylases are checkpoints in adipocyte differentiation. Experimental Biology Meeting; Boston, MA, April 21, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Kelly AM, Han ZJ, Zempleni J, Pannier AK. Enhancing Nonviral Gene Delivery to Human Mesenchymal Stem Cells through Upregulation of the Glucocorticoid Receptor. Biomedical Engineering Society Annual Meeting, Seattle, WA, September 26-28, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zempleni J. Epigenetic mechanisms of gene regulation by holocarboxylase synthetase. Invited presentation at the symposium titled Nutrigenomics and Personalized Foods held by the Korea Food Research Institute, Seoul, South Korea, October 25th, 2013.


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: I have reached my target audience using the following mechanisms: (1) publications in peer-reviewed journals; (2) presentations at local, national, and international conferences; (3) website. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Four postdoctoral fellows, seven graduate students, and 5 undergraduate students have been trained. How have the results been disseminated to communities of interest? Ten peer-reviewed papers have been published and submitted. Research findings from the project have been disseminated through 15 presentations at the local, national, and international level. Biosketches and research interests and findings of multistate group investigators are displayed on the nutrigenomics.unl.edu website in Nebraska. What do you plan to do during the next reporting period to accomplish the goals? This is the final report for W-2002. In the project that was re-authorized (W-3002), we will expand the research in cell signaling and gene regulation to (1) include food-borne microRNAs with a particular focus on milk and bone health, (2) assess natural and synthetic compounds that alter the anchoring of acetyl-CoA carboxylase in the outer mitochondrial membrane, (3) characterize physical interactions among HLCS, histone deacetylases, and the nuclear co-repressor N-CoR, and (4) assess the role of the biotin in activation of the transcription factor and tumor suppressor MBP-1.

Impacts
What was accomplished under these goals? We have created numerous antibodies against biotinylated histones. We have shown that the roles of biotin in gene repression are due to physical interactions between holocarboxylase synthetase (HLCS) and other chromatin proteins and that histone biotinylation sites are mere marks for HLCS docking sites in human chromatin. We demonstrated that biotin, methyl donors, HLCS, and chromatin proteins synergize in the repression of repeats in mammalian chromatin, thereby contributing toward genome stability and low cancer risk. This project included collaborations with more than 10 scientists that can be linked to publications, in addition to numerous other collaborations.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Eng WK, Giraud D, Schlegel VL, Wang D, Lee BH, Zempleni J. Identification and assessment of markers of biotin status in healthy adults. Br J Nutr 110:321-329
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Singh MP, Wijeratne SSK, Zempleni J. Biotinylation of lysine 16 in histone H4 contributes toward nucleosome condensation. Arch Biochem Biophys 529:105-111
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Camara Teixeira D, Malkaram SA, Zempleni J. Enrichment of meiotic recombination hotspot sequences by avidin capture technology. Gene 516:101-106
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xue J, Wijeratne SSK, Zempleni J. Holocarboxylase synthetase synergizes with methyl CpG binding protein 2 and DNA methyl transferase 1 in the transcriptional repression of long terminal repeats. Epigenetics 8:504-511
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Li Y, Hassan YI, Moriyama H, Zempleni J. Holocarboxylase synthetase interacts physically with euchromatic histone-lysine N-methyltransferase, linking histone biotinylation with methylation events. J Nutr Biochem 24:1446-1452
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xia M, Malkaram SA, Zempleni J. Three promoters regulate the transcriptional activity of the human holocarboxylase synthetase gene. J. Nutr Biochem 24: 1963-1969
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Xue J, Zhou J, Zempleni J. Holocaboxylase synthetase catalyzes biotinylation of heat shock protein 72, thereby inducing RANTES expression in HEK293 cells. Am J Physiol Cell Physiol
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Xue J, Zempleni J. Epigenetic synergies among biotin, folate, and holocarboxylase synthetase in the regulation of pro-inflammatory cytokines and repeats. Scand J Immunol 78: 419-425
  • Type: Journal Articles Status: Under Review Year Published: 2013 Citation: Baier SR, Zbasnik R, Schlegel V, Zempleni J. Off target effects of sulforaphane include the de-repression of long-terminal repeats through histone acetylation events
  • Type: Journal Articles Status: Under Review Year Published: 2013 Citation: Li Y, Malkaram SA, Zhou J, Zempleni J. Lysine biotinylation and methionine oxidation in the heat shock protein HSP60 synergize in the elimination of reactive oxygen species in human cell cultures
  • Type: Book Chapters Status: Published Year Published: 2012 Citation: Zempleni J, Wijeratne SSK, Kuroishi T. Chapter 23: Biotin. In: Present Knowledge in Nutrition. Erdman JW Jr (ed.), 10th edition. International Life Sciences Institute, Washington, DC, pp. 587-609
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Zempleni J, Liu D, Xue J. Nutrition, histone epigenetic marks, and disease. In: Epigenomics in Health and Disease. Jirtle, RL, Tyson F (eds.), Springer, Heidelberg, Germany, pp. 197-217
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Zempleni J, Liu D, Camara Teixeira D, Singh MP. Mechanisms of gene transcriptional regulation through biotin and biotin-binding proteins in mammals. In: Vitamin-binding Proteins  Their Functional Consequences. Dakshinamurti D, Dakshinamurti S (eds.), Taylor & Francis, Boca Raton, FL, pp. 219-228
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Zempleni J, Cordonier EL, Baier SR, Xue J. Vitamins, Bioactive Food Compounds, and Histone Modifications. In: Handbook of Vitamins. Zempleni J, Suttie JW, Gregory JF III, Stover PJ (eds), 5th edition. Taylor and Francis, Inc., Boca Raton, FL
  • Type: Book Chapters Status: Under Review Year Published: 2013 Citation: Zempleni J, Barshop B, Cordonier EL, Baier SR. Disorders of biotin metabolism. In: The Online Metabolic & Molecular Bases of Inherited Disease. Editors: Valle, Baudet, Vogelstein, Kinzler, Antonarakis, Ballabio, Scriver (Emeritus), Childs (Emeritus), Sly (Emeritus), Bunz (Parts Editor), Gibson (Parts Editor), Mitchell (Parts Editor). McGraw Hill.
  • Type: Journal Articles Status: Under Review Year Published: 2013 Citation: Zempleni J, Liu D. Teixeira Camara D, Cordonier EL. Novel roles of holocarboxylase synthetase in gene regulation and intermediary metabolism
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Baier SR, Schlegel VL, Zempleni J. Off Target Effects of Dietary Sulforaphane. College of Education and Human Sciences Research Fair; Lincoln, NE, November 2
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Camara D, Malkaram SA, Zempleni J. Enrichment of meiotic recombination hotspot sequences by avidin capture technology. Experimental Biology Meeting; Boston, MA, April 23
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Xue J, Zempleni J. Epigenetic synergies between methyl donors and biotin in gene repression are mediated by holocarboxylase synthetase (HLCS). Experimental Biology Meeting; Boston, MA, April 23
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Baier SR. Zbasnik R, Schlegel VL, Zempleni J. Dietary sulforaphane elicits off-target effects at loci coding for long terminal repeats in lymphocytes from healthy adults and in IMR-90 fibroblast cultures, possibly impairing genome stability. Experimental Biology Meeting; Boston, MA, April 23
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhou J, Wijeratne SSK, Zempleni J. Biotinylation of the c-myc promoter binding protein MBP-1 decreases c-myc expression in mammary carcinoma MCF-7 cells. Experimental Biology Meeting; Boston, MA, April 21
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cordonier EL, Teixeira Camara D, Han Z, Pannier AK, Zempleni J. Acetyl-CoA carboxylases are checkpoints in adipocyte differentiation. Experimental Biology Meeting; Boston, MA, April 21
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Kelly AM, Han ZJ, Zempleni J, Pannier AK. Enhancing Nonviral Gene Delivery to Human Mesenchymal Stem Cells through Upregulation of the Glucocorticoid Receptor. Biomedical Engineering Society Annual Meeting, Seattle, WA, September 26-28
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Camara D, Malkaram SA, Zempleni J. Enrichment of meiotic recombination hotspot sequences by avidin capture technology. Nebraska Gateway to Nutrigenomics Retreat. Lincoln, NE, May 13
  • Type: Books Status: Published Year Published: 2013 Citation: Handbook of Vitamins. Zempleni J, Suttie JW, Gregory JF III, Stover PJ (eds), 5th edition. Taylor and Francis, Inc., Boca Raton, FL
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Cordonier EL, Teixeira Camara D, Han Z, Pannier AK, Zempleni J. Acetyl-CoA carboxylases are checkpoints in adipocyte differentiation. Nebraska Gateway to Nutrigenomics Retreat. Lincoln, NE, May 13
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Xue J, Zhou J, Wijeratne SSK, Zempleni J. Holocarboxylase synthetase catalyzes the covalent binding of biotin to lysine residues in the inducible heat shock protein 72. Nebraska Gateway to Nutrigenomics Retreat. Lincoln, NE, May 13
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Baier SR. Zbasnik R, Schlegel VL, Zempleni J. Dietary sulforaphane elicits off-target effects at loci coding for long terminal repeats in lymphocytes from healthy adults and in IMR-90 fibroblast cultures, possibly impairing genome stability. Nebraska Gateway to Nutrigenomics Retreat. Lincoln, NE, May 13
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhou J, Wijeratne SSK, Zempleni J. Biotinylation of the c-myc promoter binding protein MBP-1 decreases c-myc expression in mammary carcinoma MCF-7 cells. Nebraska Gateway to Nutrigenomics Retreat. Lincoln, NE, May 13
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zempleni J. Roles of biotin and holocarboxylase synthetase in disease prevention. W2002 Multistate Meeting at the University of Nebraska-Lincoln, Lincoln, NE, June 3
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zempleni J. Epigenetic mechanisms of gene regulation by holocarboxylase synthetase. Invited presentation at the symposium titled Nutrigneomics and Personalized Foods held by the Korea Food Research Institute, Seoul, South Korea, October 25
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Wang Q-W, Xue J, Zempleni J. Holocarboxylase Synthetase Catalyzes Biotinylation of Lysine Residues in Enolase-1. University of Nebraska-Lincoln Undergraduate Research Symposium. August 7


Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: Discoveries have been disseminated at the Experimental Biology meeting in San Diego, CA (4/2012), at the NIFA Multistate meeting in Fort Collins (6/2013), in 11 full-length research publications, 4 book chapters, 1 review article, 6 meeting presentations by my students and postdocs, and 14 invited presentations at the local and international level. One student (Wei K. Eng) has graduated with a M.S. degree and has secured a position in a dietetic intership program in Marietta, GA. She is now pursuing a professional degree as a registered dietitian. PARTICIPANTS: Zempleni lab: Wei Kay ENG, M.S. student Elizabeth Cordonier, Ph.D. student No collaborators in the W2002 group. Collaborators from outside the W2002 group include the following faculty: Pat Dussault, Chemistry Department, University of Nebraska-Lincoln Joel Eissenberg, Department of Biochemistry, St. Louis University School of Medicine Larry Harshman, School of Biological Sciences, University of Nebraska-Lincoln Donald M. Mock, M.D., Ph.D., Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences Hideaki Moriyama, Chemistry Department, University of Nebraska-Lincoln Angela Pannier, Department of Biological Systems Engineering, University of Nebraska-Lincoln Vicki Schlegel, Department of Food Science and Technology, University of Nebraska-Lincoln John West, Department of Microbiology and Immunology at the University of Oklahoma Health Sciences Center, Oklahoma City, OK Dong Wang, Department of Statistics, University of Nebraska-Lincoln TARGET AUDIENCES: Policy makers and federal officials Faculty Industry partners Graduate and undergraduate students General public PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
One student has graduated from the program. Three postdocs, 5 doctoral students, 1 undergraduate student, and one visiting student were partially supported through this grant. The peer recognition of NIFA and the Agricultural Research Station at the University of Nebraska-Lincoln have been increased through publications in journals and books, and through seminars.

Publications

  • Liu D, Zempleni J. Holocarboxylase synthetase (HLCS) interacts physically with nuclear corepressor (N-CoR) and histone deacetylases (HDACs) to mediate gene repression. Oral presenttation in minisymposium "Nutrition and Epigenetics" (Chairs: Zempleni J, Ross SA), Experimental Biology Meeting; San Diego, CA, 8:45 - 9:00 a.m., April 22, 2012
  • Cordonier EL, Kasputis T, Mills JD, Han Z, Pannier AK, Zempleni J. Changes in the carboxylase profile are associated with early and late differentiation stages of osteoblast and adipocytes from human mesenchymal stem cells. Abstract C153 (1018.1) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 24, 2012
  • Eng WK, Schlegel VL, Wang D, Zempleni J. Development of an outpatient biotin feeding protocol for studies of biotin requirements in adults. Abstract C155 (1018.3) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 24, 2012
  • Singh D, Pannier AK,* Zempleni J. Identification of holocarboxylase synthetase chromatin binding sites using the DamID technology. Anal Biochem 413:55-59, 2011
  • Bao B, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. Human holocarboxylase synthetase with a start site at methionine-58 is the predominant nuclear variant of this protein and has catalytic activity. Biochem Biophys Res Commun 412:115-120, 2011
  • Zhou J, Wang D, Schlegel V, and Zempleni J. Development of an internet based system for modeling biotin metabolism using Bayesian networks. Comp Methods Progr Biomed 104:254-259, 2011
  • Kuroishi T, Rios-Avila L, Pestinger V, Wijeratne SSK, Zempleni J. Biotinylation is a natural, albeit rare, modification of human histones Mol Genet Metabol 104:537-545, 2011
  • Esaki S, Malkaram SA, Zempleni J. Effects of single nucleotide polymorphisms in the human holocarboxylase synthetase gene on enzyme catalysis. Eur J Hum Genet 20:428-433, 2012
  • Rios-Avila L, Pestinger V, Wijeratne SSK, Zempleni J. K16-biotinylated histone H4 is overrepresented in repeat regions and participates in the repression of transcriptionally competent genes in human Jurkat lymphoid cells. J Nutr Biochem 23:1559-1564, 2012
  • Eng WK, Giraud D, Schlegel VL, Wang D, Lee BH, Zempleni J. (2012) Identification and assessment of markers of biotin status in healthy adults. Br J Nutr (in press)
  • Singh MP, Wijeratne SSK, Zempleni J. (2012) Biotinylation of lysine 16 in histone H4 contributes toward nucleosome condensation. Arch Biochem Biophys (in press)
  • Li Y, Hassan YI, Moriyama H, Zempleni J. (2012) Holocarboxylase synthetase interacts physically with euchromatic histone-lysine N-methyltransferase, linking histone biotinylation with methylation events (submitted)
  • Camara Teixeira D, Malkaram SA, Zempleni J. (2012) Enrichment of meiotic recombination hotspot sequences by avidin capture technology (submitted)
  • Zempleni J, Liu D, Xue J. (2012) Nutrition, histone epigenetic marks, and disease. In: Epigenomics in Health and Disease. Jirtle, RL, Tyson F (eds.), Springer, Heidelberg, Germany (in press)
  • Bao B, Rodriguez-Melendez R, Zempleni J. Cytosine methylation in miR-153 gene promoters increases the expression of holocarboxylase synthetase, thereby increasing the abundance of histone H4 biotinylation marks in HEK-293 human kidney cells. J Nutr Biochem 23:635-639, 2011
  • Zempleni, J, Eng WK, Singh MP, Baier SR. The chemistry and biochemistry of biotin. In: Food and Nutritional Components in Focus. Preedy VR (ed.), Royal Society of Chemistry, London, U.K., 2012, pp. 146-157
  • Zempleni J, Liu D, Camara Teixeira D, Singh MP. (2012) Mechanisms of gene transcriptional regulation through biotin and biotin-binding proteins in mammals. In: Vitamin-binding Proteins - Their Functional Consequences. Dakshinamurti D, Dakshinamurti S (eds.), Taylor & Francis, Boca Raton, FL (invited manuscript, in press)
  • Zempleni J, Cordonier EL, Baier SR, Xue J. (2012) Vitamins, Bioactive Food Compounds, and Histone Modifications. In: Handbook of Vitamins. Zempleni J, Suttie JW, Gregory JF III, Stover PJ (eds), 4th edition. Taylor and Francis, Inc., Boca Raton, FL, 2013 (submitted)
  • Malkaram SA, Hassan YI, Zempleni J. Online tools for bioinformatics analyses in nutrition sciences. Adv Nutr 3:654-665, 2012
  • Xia M, Malkaram SA, Zempleni J. Identification of three promoters in the human holocarboxylase synthetase (HCS) gene. Abstract 416/C298 (647.1) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 22, 2012
  • Wijeratne SSK, Malkaram SA, Pabian MD, Granatowicz AD, Zempleni J. Identification of biotin- and holocarboxylase synthetase-dependent microRNAs in human fibroblasts. Abstract C300 (647.3) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 22, 2012
  • Pratap Singh M, Zempleni J. Biotinylation of K16 in histone H4 causes chromatin condensation. Oral presenttation in minisymposium "Nutrition and Epigenetics" (Chairs: Zempleni J, Ross SA), Experimental Biology Meeting; San Diego, CA, 9:00 - 9:15 a.m., April 22, 2012


Progress 10/01/10 to 09/30/11

Outputs
OUTPUTS: (1) Results have been disseminated at national meetings, e.g., Experimental Biology. In addition, seminars have been presented at various research institutions and to the lay public in the state of Nebraska and at both the national and international level. (2) Unique reagents have been shared free of charge with qualified investigators in the U.S. (3) A student has graduated with M.S. degrees from the University of Nebraska-Lincoln. PARTICIPANTS: Subhashinee Wijeratne (UNL), Rocio Rodriguez-Melendez (UNL), Baolong Bao (UNL), Zhongji Han (UNL), Shingo Esaki (UNL), Jing Xue (UNL), Dandan Liu (UNL), Scott Baier (UNL), Elizabeth Cordonier (UNL), Daniel Camara Teixeira (UNL), Wei Kay Eng (UNL), Luisa Rios (UNL), Valerie Pestinger (UNL), Toshinobu Kuroishi (UNL), Mahendra Pratap Singh (UNL), Yong Li (UNL), David Giraud (UNL), Gaganpreet Kaur Mall (UNL), Kate Roehrs (UNL), Andrew Granatowicz (UNL), Michael Pabian (UNL), Angela Pannier (UNL), Don Mock (Little Rock, AR), John West (Oklahoma), Joel Eissenberg (Saint Louis), Yie-Hwa Chang (St. Louis), Dong Wang (UNL), Vicki Schlegel (UNL), Gloria Borgstahl (UNMC), Yuri Lyubchenko (UNMC). TARGET AUDIENCES: Scientists, policy makers, graduate and undergraduate students, publishers, funding agencies, lay public. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Outcomes: (1) Resources from this project paid for supplies to complete one M.S. thesis and continues to support four postdoctoral fellows, four Ph.D. students, one M.S. student, and two technicians. (2) Resources from this project helped to generate numerous publications as described below. Impacts: (1) Developed workforce in the molecular sciences area. (2) Enhanced recognition by peer institutions. (3) Enhanced the competitiveness for additional external funding. (4) Increased the scientific knowledge base in nutrition.

Publications

  • Pestinger V, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. The histone biotinylation marks H3K9bio, H3K18bio, and H4K8bio are enriched in repeat regions and participate in the repression of transcriptionally competent genes in human primary fibroblasts and Jurkat lymphoblastoma cells. J Nutr Biochem 22:328-333, 2011
  • Bao B, Pestinger V, Hassan YI, Borgstahl GEO, Kolar C, Zempleni J. Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18. J Nutr Biochem 22:470-475, 2011
  • Filenko NA, Kolar C, West JT, Hassan YI, Borgstahl GEO, Zempleni J, Lyubchenko YL. The role of histone H4 biotinylation in the structure and dynamics of nucleosomes. PLoS ONE 6:e16299, 2011
  • Rios-Avila L, Prince SA, Wijeratne SSK, Zempleni J. A 96-well plate assay for high-throughput analysis of holocarboxylase synthetase activity. Clin Chim Acta 412:735-739, 2011
  • Singh D, Pannier AK, Zempleni J. Identification of holocarboxylase synthetase chromatin binding sites using the DamID technology. Anal Biochem 413:55-59, 2011
  • Bao B, Rodriguez-Melendez R, Zempleni J. Cytosine methylation in miR-153 gene promoters increases the expression of holocarboxylase synthetase, thereby increasing the abundance of histone H4 biotinylation marks in HEK-293 human kidney cells. J Nutr Biochem (in press), 2012
  • Bao B, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. Human holocarboxylase synthetase with a start site at methionine-58 is the predominant nuclear variant of this protein and has catalytic activity. Biochemical and Biophysical Research Communications 412:115-120, 2011
  • Zhou J, Wang D, Schlegel V, and Zempleni J. Development of an internet based system for modeling biotin metabolism using Bayesian networks. Comp Methods Progr Biomed (in press), 2012
  • Kuroishi T, Rios-Avila L, Pestinger V, Wijeratne SSK, Zempleni J. Biotinylation is a natural, albeit rare, modification of human histones Mol Genet Metabol (in press), 2012
  • Esaki S, Malkaram SA, Zempleni J. Effects of single nucleotide polymorphisms in the human holocarboxylase synthetase gene on enzyme catalysis. Eur J Hum Genet (in press), 2012
  • Rios-Avila L, Pestinger V, Wijeratne SSK, Zempleni J. K16-biotinylated histone H4 is overrepresented in repeat regions and participates in the repression of transcriptionally competent genes in human Jurkat lymphoid cells. J Nutr Biochem (in press), 2012
  • Xue J, Zempleni J. Epigenetic synergies between methylation of cytosines and biotinylation of histones in gene repression. Abstract 249 (597.7) Experimental Biology Meeting; Washington, DC, April 10, 2011
  • Esaki S, Zempleni J. Effects of single nucleotide polymorphisms in the human holocarboxylase synthetase gene on catalytic activity. Abstract 291 (782.7) Experimental Biology Meeting; Washington, DC, April 11, 2011
  • Zempleni J. Enhancing nutrigenomics research in Auckland. University of Auckland, New Zealand, February 3, 2011.
  • Wijeratne SSK, Zempleni J. Identification of HCS-interacting proteins through the CytoTrapTM two-hybrid system. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Eng WK, Giraud D, Zempleni J. Development of an outpatient biotin feeding protocol for studies of biotin biology in adults. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Teixeira D, Malkaram SA, Zempleni J. Detection and enrichment of a common DNA sequence associated with human genome instability. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Malkaram SA, Wijeratne SSK, Zempleni J. High-throughput ChIP-seq and RNA-seq investigation of epigenetic regulation of gene expression by biotin. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Li Y, Zempleni J. Holocarboxylase synthetase interacts with euchromatic histone-lysine N-methyltransferase 1, linking histone biotinylation to histone methylation. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Xue J, Zempleni J. Epigenetic synergies between methylation of cytosines and biotinylation of histones in gene repression. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Xue J, Zempleni J. Effects of single nucleotide polymorphisms in the human holocarboxylase synthetase gene on catalytic activity. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Zempleni J. Biotin-dependent epigenetic mechanisms contributing to genome stability. University of Vienna, Austria, June 16, 2011.
  • Zempleni J, Li Y, Xue J, Cordonier EL. The role of holocarboxylase synthetase in genome stability is mediated partly by epigenomic synergies between methylation and biotinylation events. Epigenetics 6:892-894, 2011
  • Zempleni J, Camara Teixeira D, Kuroishi T, Cordonier EL, Baier S. Biotin requirements for DNA damage prevention. Mutat Res (in press), 2012
  • Zempleni J, Kuroishi T. Nutrition information brief - biotin. Adv Nutr 2011 (in press), 2012
  • Kuroishi T, Zempleni J. Creation of holocarboxylase synthetase knockdown murine primary fibroblasts. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.
  • Pratap Singh M, Zempleni J. Effects of biotinylation of lysine-16 in histone H4 on nucleosomal assembly. Nebraska Gateway for Nutrigenomics Retreat. Lincoln, NE, February 28th, 2011.


Progress 10/01/09 to 09/30/10

Outputs
OUTPUTS: (1) Results have been disseminated at national meetings, e.g., Experimental Biology. In addition, seminars have been presented at various research institutions and to the lay public in the state of Nebraska and nationally. (2) Unique reagents have been shared free of charge with qualified investigators in the U.S. (3) Students have graduated with M.S. degrees from the University of Nebraska-Lincoln. PARTICIPANTS: Subhashinee Wijeratne (UNL), Rocio Rodriguez-Melendez (UNL), Baolong Bao (UNL), Shingo Esaki (UNL), Jing Xue (UNL), Dandan Liu (UNL), Wei Kay Eng (UNL), Luisa Rios (UNL), Valerie Pestinger (UNL), Toshinobu Kuroishi (UNL), Mahendra Pratap Singh (UNL), Yong Li (UNL), David Giraud (UNL), Gaganpreet Kaur Mall (UNL), Kate Roehrs (UNL), Andrew Granatowicz (UNL), Michael Pabian (UNL), John West (Oklahoma), Joel Eissenberg (Saint Louis), Yie-Hwa Chang (St. Louis), Dong Wang (UNL), Vicki Schlegel (UNL), Gloria Borgstahl (UNMC), Yuri Lyubchenko (UNMC), and Michael Brattain (UNMC). TARGET AUDIENCES: Scientists, policy makers, graduate and undergraduate students, publishers and funding agencies. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Outcomes: (1) Resources from this project paid for supplies to complete three M.S. theses. (2) Resources from this project helped to generate numerous publications as described below. Impacts: (1) Developed workforce in the molecular sciences area. (2) Enhanced recognition by peer institutions. (3) Enhanced the competitiveness for additional external funding. (4) Increased the scientific knowledge base in nutrition.

Publications

  • Wijeratne SSK, Camporeale G, Zempleni J. K12-biotinylated histone H4 is enriched in telomeric repeats from human lung IMR-90 fibroblasts. J Nutr Biochem 21:310-316, 2010
  • Hassan YI, Moriyama H, Zempleni J. The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. Arch Biochem Biophys 495:35-41, 2010
  • Kaur Mall G, Chew YC, Zempleni J. The mechanisms of biotin homeostasis are qualitatively similar but quantitatively different in human Jurkat lymphoid and HepG2 liver cells. J Nutr 140:1086-1092, 2010
  • Bao B, Rodriguez-Melendez R, Wijeratne SSK, Zempleni J. Biotin regulates the expression of holocarboxylase synthetase in a miR-539 pathway in HEK-293 human embryonic kidney cells. J Nutr 140:1546-1551, 2010
  • Pestinger V, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. (2010) The histone biotinylation marks H3K9bio, H3K18bio, and H4K8bio are enriched in repeat regions and participate in the repression of transcriptionally competent genes in human primary fibroblasts and Jurkat lymphoblastoma cells. J Nutr Biochem (in press)
  • Bao B, Pestinger V, Hassan YI, Borgstahl GEO, Kolar C, Zempleni J. (2010) Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18. J Nutr Biochem (in press)
  • Rios-Avila L, Pestinger V, Zempleni J. (2010) K16-biotinylated histone H4 is overrepresented in repeat regions and participates in the repression of transcriptionally competent genes in human Jurkat lymphoid cells. (submitted)
  • Rios-Avila L, Prince SA, Wijeratne SSK, Zempleni J. (2010) A 96-well plate assay for high-throughput analysis of holocarboxylase synthetase activity. (submitted)
  • Filenko NA, Kolar C, West JT, Hassan YI, Borgstahl GEO, Zempleni J, Lyubchenko YL. (2010) The role of histone H4 biotinylation in the structure and dynamics of nucleosomes (submitted)
  • Eissenberg JC, Zempleni J. Biotinylated histone isoforms have dispersed and distinct distributions in the Drosophila genome (2009, submitted)
  • Wijeratne SSK, Kuroishi T, Pestinger V, Rios L, Zempleni J. Histone biotinylation is a naturally occurring phenomenon. Abstract 2316 (107.1) Experimental Biology Meeting; Anaheim, CA, 3:00 p.m., April 25, 2010
  • Rios-Avila L, Wijeratne SSK, Pestinger V, Zempleni J. Characterization of the H4K16bio mark in human lymphoid cells. Abstract 2273 (550.1) Experimental Biology Meeting; Anaheim, CA, 12:45 - 1:45 p.m., April 25, 2010
  • Rodriguez-Melendez R, Bui DC, Ellis M, Johnson RM, Zempleni J. Identification of a potential role of K9-biotinylated histone H3 in honeybee (Apis mellifera) development. Abstract 3220 (550.2) Experimental Biology Meeting; Anaheim, CA, 1:45 - 2:45 p.m., April 25, 2010
  • Kuroishi T, Cerny RL, Zempleni J. Mass spectrometric analysis of biotinylated peptides and histones. Abstract 3809 (107.2) Experimental Biology Meeting; Anaheim, CA, 3:30 p.m., April 25, 2010
  • Singh D, Zempleni J, Pannier A. Development of an Antibody Independent Technology to Monitor Chromatin Proteins in Cell Lines. University of Nebraska Medical Center, Regenerative Medicine Symposium, May 24, 2010, Omaha, NE
  • Rios-Avila L, Pestinger V, Wijeratne SSK, Rodriguez-Melendez R, Zempleni J. Characterization of the H4K16bio mark in human cells. UNL Research Fair, April 7, 2010, Lincoln, NE
  • Hassan YI, Zempleni J. Molecular Aspects of Physical Performance and Nutritional Assessment. In: Nutritional Assessment of Athletes. Driskell JA, Wolinksy I (eds.). Taylor and Francis, Inc., Boca Raton, FL, 2010, 2nd edition: 212-232
  • Zempleni J, Wijeratne SSK, Kuroishi T. Biotin. In: Present Knowledge in Nutrition. Erdman J, Macdonald I, Zeisel S (eds.), 10th edition. International Life Sciences Institute, Washington, DC, 2012 (invited paper, submitted)
  • Ho E, Zempleni J. Overview to symposium "Nutrients and Epigenetic Regulation of Gene Expression." J Nutr 139:2387-2388, 2009
  • Zempleni J, Chew YC, Bao B, Pestinger V, Wijeratne SSK. Repression of transposable elements by histone biotinylation. J Nutr 139:2389-2392, 2009


Progress 10/01/08 to 09/30/09

Outputs
OUTPUTS: (1) Results have been disseminated at national meetings, e.g., Experimental Biology, FASEB Summer Conference, and an Epigenetics conference sponsored by the European Molecular Biology Organization. In addition seminars have been presented at various institutions in the state of Nebraska and nationally. (2) Unique reagents have been shared free of charge with qualified investigators from within the U.S. PARTICIPANTS: Subhashinee Wijeratne (UNL), Rocio Rodriguez-Melendez (UNL), Baolong Bao (UNL), Shingo Esaki (UNL), Jing Xue (UNL), Yousef Hassan (UNL), Keyna Kobza (UNL), Luisa Rios (UNL), Valerie Pestinger (UNL), Toshinobu Kuroishi (UNL), Azusa Kuroishi (UNL), David Giraud (UNL), Gaganpreet Kaur Mall (UNL), Kate Roehrs (UNL), Jenna Rickstrew (UNL), , John West (Oklahoma), Joel Eissenberg (Saint Louis), Yie-Hwa Chang (St. Louis), Dong Wang (UNL), Vicki Schlegel (UNL), Gloria Borgstahl (UNMC), Yuri Lyubchenko (UNMC), and Judith Christman (UNMC). TARGET AUDIENCES: Scientists, policy makers, graduate and undergraduate students, publishers and funding agencies. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
(1) A patent application is pending and licensing agreements with industry are in place. (2) Resources from this project paid for a M.S. student. (3) Resources from this project helped to generate numerous publications as described below.

Publications

  • Kobza KA, Chaiseeda K, Sarath G, Takacs JM, Zempleni J. Biotinyl-methyl 4-(amidomethyl) benzoate is a competitive inhibitor of human biotinidase. J Nutr Biochem 19:826-832, 2008
  • Chew YC, West JT, Kratzer SJ, Ilvarsonn AM, Eissenberg JC, Dave BJ, Klinkebiel D, Christman JK, Zempleni J. Biotinylation of histones represses transposable elements in human and mouse cells and cell lines, and in Drosophila melanogaster. J Nutr 138:2316-2322, 2008
  • Rodriguez-Melendez R, Zempleni J. Nitric oxide signaling depends on biotin in Jurkat human lymphoma cells. J Nutr 139:429-433, 2009
  • Hassan YI, Moriyama H, Olsen LJ, Bi X, Zempleni J. N- and C-terminal domains in human holocarboxylase synthetase participate in substrate recognition. Mol Genet Metabol 96:183-188, 2009
  • Wijeratne SSK, Camporeale G, Zempleni J. (2009) K12-biotinylated histone H4 is enriched in telomeric repeats from human lung IMR-90 fibroblasts. J Nutr Biochem (in press)
  • Eissenberg JC, Zempleni J. Biotinylated histone isoforms have dispersed and distinct distributions in the Drosophila genome (2009, submitted)
  • Bao B, Pestinger V, Hassan YI, Borgstahl GEO, Kolar C, Zempleni J. Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18 (2009, submitted)
  • Hassan YI, Moriyama H, Zempleni J. The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. (2009, submitted) Filenko N, Kolar C, Brodie R, Hassan YI, West JT, Borgstahl GEO, Zempleni J, Lyubchenko YL. Comparison of nucleosomes reconstituted with native histone H4 and biotinylated H4 K12C mutant. Presented at the Structural Biology and Molecular Biophysics Workshop on July 13, 2009, at the Durham Research Center at the University of Nebraska Medical Center, Omaha, NE
  • Singh D, Pannier A, Zempleni J. Developing an Antibody-Independent Technology to Monitor Chromatin Proteins in Human Breast and Human Breast Cancer Cell Lines to Map Epigenetic Profiles. NSF EPSCoR Research Conference, September 29, 2009, Omaha, NE.
  • Rios-Avila L, Zempleni J. Characterization of the H4K16bio mark in human cells. NSF EPSCoR Research Conference, September 29, 2009, Omaha, NE
  • Singh D, Zempleni J, Pannier A. Development of technologies to monitor chromatin proteins in small cell numbers for applications in assisted reproductive physiology. Biotechnology and Bioinformatics Symposium, October 9 - 11, 2009; Lincoln, NE.
  • Wijeratne SSK, Kuroishi T, Pestinger V, Rios L, Zempleni J. Histone biotinylation is a naturally occurring phenomenon. Meeting abstract submitted for the Experimental Biology Meeting; Anaheim, CA, April, 2010
  • Rios-Avila L, Wijeratne SSK, Pestinger V, Zempleni J. Characterization of the H4K16bio mark in human lymphoid cells. Meeting abstract submitted for the Experimental Biology Meeting; Anaheim, CA, April, 2010
  • Rodriguez-Melendez R, Bui DC, Ellis M, Johnson RM, Zempleni J. Identification of a potential role of K9-biotinylated histone H3 in honeybee (Apis mellifera) development. Meeting abstract submitted for the Experimental Biology Meeting; Anaheim, CA, April, 2010
  • Kuroishi T, Cerny RL, Zempleni J. Mass spectrometric analysis of biotinylated peptides and histones. Meeting abstract submitted for the Experimental Biology Meeting; Anaheim, CA, April, 2010
  • Hassan YI, Zempleni J. A novel, enigmatic histone modification: biotinylation of histones by holocarboxylase synthetase. Nutr Rev 66(12):721-725, 2008
  • Wang D, Schlegel V, Zempleni J. The use of Bayesian networks for predicting nutrient intake, metabolism, and requirements in silico. Nutr Rev (2009, invited manuscript, submitted)
  • Ho E, Zempleni J. Overview to symposium "Nutrients and Epigenetic Regulation of Gene Expression." J Nutr (2009, in press)
  • Zempleni J, Chew YC, Bao B, Pestinger V, Wijeratne SSK. Repression of transposable elements by histone biotinylation. J Nutr (2009, in press)
  • Zempleni J, Wijeratne SSK, Hassan YI. Biotin. BioFactors 35:36-46, 2009