Progress 09/01/06 to 08/31/10
Outputs
(N/A)
Impacts
One postdoctoral fellow, one technician, and four undergraduate students acquired expertise with molecular biology techniques during the final year of this project. Techniques included chromatin immunoprecipitation assays, recombinant proteins, avidin/biotin technology, statistical methods, and others. This has a positive impact on the workforce development in the state of Nebraska and the U.S.
Publications
- 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
- 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
- Hassan YI, Moriyama H, Zempleni J. (2010) The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. Arch Biochem Biophys 495:35-41
- 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)
- 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
- 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)
- 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)
- Kaur Mall G, Chew YC, Zempleni J. (2010) The mechanisms of biotin homeostasis are qualitatively similar but quantitatively different in human Jurkat lymphoid and HepG2 liver cells. J Nutr 140:1086-1092
- Bao B, Rodriguez-Melendez R, Wijeratne SSK, Zempleni J. (2010) Biotin regulates the expression of holocarboxylase synthetase in a miR-539 pathway in HEK-293 human embryonic kidney cells. J Nutr 140:1546-1551
- 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
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Progress 09/01/08 to 08/31/09
Outputs
(N/A)
Impacts
Knowledge of project participants (1 postdoc, 1 technician, 2 undergraduates) changed by acquiring expertise with molecular biology techniques during year 3 of this project, e.g., chromatin immunoprecipitation assays, recombinant proteins, avidin/biotin technology, statistical methods, and others. This has a positive impact on the workforce development in the state of Nebraska and the U.S.
Publications
- Chew YC, West JT, Kratzer SJ, Ilvarsonn AM, Eissenberg JC, Dave BJ, Klinkebiel D, Christman JK, Zempleni J. (2008) Biotinylation of histones represses transposable elements in human and mouse cells and cell lines, and in Drosophila melanogaster. J Nutr 138:2316-2322
- Rodriguez-Melendez R, Zempleni J. (2009) Nitric oxide signaling depends on biotin in Jurkat human lymphoma cells. J Nutr 139:429-433
- Hassan YI, Moriyama H, Olsen LJ, Bi X, Zempleni J. (2009) N- and C-terminal domains in human holocarboxylase synthetase participate in substrate recognition. Mol Genet Metabol 96:183-188
- Wijeratne SSK, Camporeale G, Zempleni J. K12-biotinylated histone H4 is enriched in telomeric repeats from human lung IMR-90 fibroblasts. (2010) J Nutr Biochem (in press)
- Eissenberg JC, Zempleni J. (2009) Biotinylated histone isoforms have dispersed and distinct distributions in the Drosophila genome (submitted)
- Bao B, Pestinger V, Hassan YI, Borgstahl GEO, Kolar C, Zempleni J. (2009)Holocarboxylase synthetase is a chromatin protein and interacts directly with histone H3 to mediate biotinylation of K9 and K18 (submitted)
- Hassan YI, Moriyama H, Zempleni J. (2009)The polypeptide Syn67 interacts physically with human holocarboxylase synthetase, but is not a target for biotinylation. (submitted)
- Wijeratne SSK, Zempleni J. (2009) K12-biotinylated histone H4 is enriched in human telomeric repeats. Abstract 1246 (B192) Experimental Biology Meeting; New Orleans, LA, 1:45 - 2:45 p.m., April 19
- Bao B, Pestinger V, Zempleni J. (2009) Holocarboxylase synthetase physically interacts with histone H3 to mediate biotinylation of K9 and K18. Abstract 1414 (B194); Experimental Biology Meeting; New Orleans, LA, 12:45 - 1:45 p.m., April 19
- Pestinger V, Wijeratne SSK, Zempleni J. (2009) Enrichment of H3K4bio, H3K9bio, H3K18bio, and H4K8bio in distinct genomic loci. Abstract 1296 (B193); Experimental Biology Meeting; New Orleans, LA, 1:45 - 2:45 p.m., April 19
- Mall GK, Chew YC, Zempleni J. (2009) Homeostasis of biotin in human lymphoma and liver cells. Abstract 1381 (B142) Experimental Biology Meeting; New Orleans, LA, 12:45 - 1:45 p.m., April 19
- Rodriguez-Melendez, Zempleni J. (2009) Nitric oxide signaling depends on biotin in Jurkat human lymphoma cells. Abstract 1711 (B124); Experimental Biology Meeting; New Orleans, LA, 12:45 - 1:45 p.m., April 19
- Filenko N, Kolar C, Brodie R, Hassan YI, West JT, Borgstahl GEO, Zempleni J, Lyubchenko YL. (2009) 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 at the Durham Research Center at the University of Nebraska Medical Center, Omaha, NE
- Singh D, Pannier A*, Zempleni J*. (2009) 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, Omaha, NE. *These authors contributed equally to the project.
- Rios-Avila L, Zempleni J. (2009) Characterization of the H4K16bio mark in human cells. NSF EPSCoR Research Conference, September 29, Omaha, NE.
- Rodriguez-Melendez R, Hassan YI, Zempleni J. (2009) Mechanisms of regulation of the interleukin-2 gene by biotin. Presented in conjunction with the IFT (Institute of Food Technologists) meeting on June 6 in Anaheim, CA
- Singh D, Zempleni J*, Pannier A*. (2009) Development of technologies to monitor chromatin proteins in small cell numbers for applications in assisted reproductive physiology. Biotechnology and Bioinformatics Symposium, October 9 - 11; Lincoln, NE. *These authors contributed equally to the project.
- Zempleni J, Wijeratne SSK, Hassan YI. (2009) Biotin. BioFactors 35:36-46
- Wang D, Schlegel V, Zempleni J. (2009) The use of Bayesian networks for predicting nutrient intake, metabolism, and requirements in silico. Nutr Rev (invited manuscript, submitted)
- Zempleni J, Gralla M, Camporeale G, Hassan YI. (2009) The sodium-dependent multivitamin transporter (SMVT) gene is regulated at the chromatin level by histone biotinylation in human Jurkat lymphoblastoma cells. J Nutr 139:163-166
- Ho E, Zempleni J. (2009) Overview to symposium "Nutrients and Epigenetic Regulation of Gene Expression." J Nutr (in press)
- Zempleni J, Chew YC, Bao B, Pestinger V, Wijeratne SSK. (2009) Repression of transposable elements by histone biotinylation. J Nutr (in press)
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Progress 09/01/07 to 08/31/08
Outputs
(N/A)
Impacts
Knowledge of project participants (1 postdoc, two undergraduates)changed by acquiring expertise with molecular biology techniques during year 2 of this project, e.g., chromatin immunoprecipitation assays, recombinant proteins, avidin/biotin technology, statistical methods, and others. Positive life choices were made by directing the two undergraduate students towards graduate programs in nutrition and health sciences, and pharmacy. This also positively impacted the workforce development in the state of Nebraska and the U.S.
Publications
- Camporeale G. Oommen AM, Griffin JB, Sarath G, Zempleni J. (2007) K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells. J Nutr Biochem 18:760-768
- Gralla M, Camporeale G, Zempleni J. 2008. Holocarboxylase synthetase regulates expression of biotin transporters by chromatin remodeling events at the SMVT locus. J Nutr biochem 19:400-408.
- Kobza KA, Chaiseeda K, Sarath G, Takacs JM, Zempleni J. 2008. Biotinyl-methyl 4-(amidomethyl) benzoate is a competitive inhibitor of human biotinidase. J Nutr Biochem. (in press).
- Chew YC, West JT, Kratzer SJ, Ilvarsonn AM, Eissenberg JC, Dave BJ, Klinkebiel D, Christman JK, Zempleni J. 2008. A diet-dependent epigenetic mechanism that represses transposable elements (submitted)
- Wijeratne SSK, Camporeale G, Zempleni J. 2008. K12-biotinylated histone H4 is enriched in telomeric repeats from human lung IMR-90 fibroblasts (submitted)
- Hassan YI, Zempleni J. (2007) A tyrosine kinase is involved in the nuclear translocation of human holocarboxylase synthetase and, hence, binding of the vitamin biotin to histones. Presented by Yi Hassan at the AICR/WCRF International Research Conference on Food, Nutrition and Cancer in Washington DC (November 1 and 2, 2007)
- Chew YC, West JT, Kratzer SJ, Ilvarsonn AM, Eissenberg JC, Zempleni J. 2008. Histone biotinylation represses retrotransposons in whole organisms, decreasing production of viral particles and retrotranspositions. Abstract C119 (689.1) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 6, 2008
- Wijeratne SSK, Zempleni J. Biotinylation and methylation of histones play a role in regulation of genes associated with hydrogen peroxide induced oxidative stress. 2008. Abstract C119 (689.2) Experimental Biology Meeting; San Diego, CA, 1:45 - 2:45 p.m., April 6, 2008
- Rodriguez-Melendez R, White B, Zempleni J. 2008. Biotin-dependent cell signaling networks in mouse embryos. Abstract C119 (689.3) Experimental Biology Meeting; San Diego, CA, 12:45 - 1:45 p.m., April 6, 2008
- Hassan YI, Zempleni J. 2008. A tyrosine kinase is involved in the nuclear translocation of human holocarboxylase synthetase. Abstract C119 (691.14) Experimental Biology Meeting; San Diego, CA, 1:45 - 2:45 p.m., April 6, 2008
- Kobza K, Sarath G, Zempleni J. 2008. Prokaryotic BirA ligase biotinylates K4, K9, K18 and K23 in histone H3. Biochemistry and Molecular Biology Reports (BMB Reports) 41:310-315.
- Zempleni J, Chew YC, Hassan YI, Wijeratne SSK. 2008. Epigenetic regulation of chromatin structure and gene function by biotin: are biotin requirements being met Nutr Rev (in press)
- Zempleni J, Gralla M, Camporeale G, Hassan YI. 2008. The sodium-dependent multivitamin transporter (SMVT) gene is regulated at the chromatin level by histone biotinylation in human Jurkat lymphoblastoma cells. J Nutr (in press)
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Progress 09/01/06 to 08/31/07
Outputs
The long-term goal of this project is to identify biotin-dependent mechanisms that affect human immune function. The following major observations have been made. (1) Biotin deficiency impairs immune function (mediated by interleukin-2) due to decreased expression of the interleukin-2 receptor gamma gene. The high concentrations of interleukin-2 observed in biotin-deficient cell cultures are artifacts, caused by decreased receptor-mediated endocytosis of interleukin-2. In fact, the transcription of the interleukin-2 gene is decreased in biotin-deficient cells compared with normal controls. (2) We have identified a number of signaling pathways that mediated decreased expression of interleukin-2 and interleukin-2 receptor gamma in biotin-deficient human immune cells. These pathways include NF-kB, AP1, and perhaps nitric oxide. (3) A biotin-responsive element has been identified in the region approximately 1000 basepairs upstream from the transcription start site of the
interleukin-2 gene. (4) We have demonstrated that binding of biotin to lysine-12 in histone H4 plays a critical role in regulating the transcriptional activity of the human interleukin-2 gene. An assay for histone debiotinylases has been developed. (5) Decreased biotinylation of histones is associated with increased susceptibility to heat stress in eukaryotes. (6) Proteins that interact with the histone biotinyl transferase Holocarboxylase Synthetase have been tentatively identified. (7) Decreased histone biotinylation at endogenous retroviral elements impairs chromosomal stability.
Impacts
Biotin deficiency is fairly common in the U.S. population. For example, evidence has been provided that 50% of pregnant women are marginally biotin deficient. Similar scenarios can be found in individuals treated with anticonvulsants. Our research suggests that biotin-deficient individuals are at increased risk for being immune deficient. Our research offers mechanistic insights in how the nutrient biotin affects immune function. It is reasonable to assume that biotin supplementation has beneficial effects on immune function in populations at risk, e.g., pregnant women and patients treated with anticonvulsants.
Publications
- Camporeale G, Zempleni J, Eissenberg JC. (2007) Susceptibility to heat stress and aberrant gene expression patterns in holocarboxylase synthetase-deficient Drosophila melanogaster are caused by decreased biotinylation of histones, not of carboxylases. J Nutr 137:885-889.
- Chew YC, Sarath G, Zempleni J. (2007) An avidin-based assay for histone debiotinylase activity in human cell nuclei. J Nutr Biochem 18:475-481.
- Smith EM, Hoi JT, Eissenberg JC, Shoemaker JD, Neckameyer WS, Ilvarsonn AM, Harshman LG, Schlegel VL, Zempleni J. (2007) Feeding Drosophila a biotin-deficient diet for multiple generations increases stress resistance and lifespan and alters gene expression and histone biotinylation patterns. J Nutr 137:2006-2012.
- Gralla M, Camporeale G, Zempleni J. (2007) Holocarboxylase synthetase regulates expression of biotin transporters by chromatin remodeling events at the SMVT locus. J Nutr Biochem (in press).
- Kobza KA, Chaiseeda K, Sarath G, Takacs JM, Zempleni J. (2007) Biotinyl-methyl 4-(amidomethyl) benzoate is a competitive inhibitor of human biotinidase (submitted).
- Smith EM, Zempleni J. (2007) Protein biotinylation signatures in malignant and proliferating cells. (submitted).
- Zempleni J, Chew YC, Hassan YI, Wijeratne, SSK. (2008) Epigenetic regulation of chromatin structure and gene function by biotin: are biotin requirements being met? Nutr Rev (invited manuscript in preparation).
- Wijeratne SSK, Zempleni J. (2007) Decreased histone biotinylation marks cells senescence. Abstract D286 (855.2) Experimental Biology Meeting; Washington, D.C., 1:45 - 2:45 p.m., May 1.
- Camporeale G, Oommen AM, Griffin JB, Sarath G, Zempleni J. (2007) K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells. J Nutr Biochem (in press).
- Chew YC, West JT, Zempleni J. (2007) K12 biotinylated histone H4 is enriched at human endogenous retrovirus promoter regions and may function in retroviral silencing. Abstract D287 (855.3) Experimental Biology Meeting; Washington, D.C., 12:45 - 1:45 p.m., May 1.
- Hassan YI, Atkin A, Zempleni J. (2007) Protein-protein interactions of human holocarboxylase synthetase reveals potential association with zinc-finger proteins. Abstract D285 (855.1) Experimental Biology Meeting; Washington, D.C., 12:45 - 1:45 p.m., May 1.
- Chew YC, West JT, Zempleni J. (2007) K12 biotinylated histone H4 is enriched at human endogenous retrovirus promoter regions and may function in retroviral silencing. Presented at the 2007 Meeting of the Microbiology Initiative at the University of Nebraska-Lincoln, August 20.
- Hassan YI, Zempleni J. (2007) A tyrosine kinase is involved in the nuclear translocation of human holocarboxylase synthetase. Presented at the 2007 Meeting of the Microbiology Initiative at the University of Nebraska-Lincoln, August 20.
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