Progress 10/01/10 to 09/30/17
Outputs
Target Audience:The findings from our research are of great interest to the public, scientists studying infection biology, research extension specialists attempting to prevent bovine mastitis,and physicians treating S. aureus infections. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This past year i have trained one undergraduate and three graduate students by conducting research on this project. Each of these students presented their work at scientific meetings. The work of the three graduate students was published in peer-reviewed journals. How have the results been disseminated to communities of interest?The results have been dissiminated through peer-review publications and paper and poster presentations at at local, national, and international scientific meetings. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
OBJECTIVE 1: Characterization of host mechanisms associated with mastitis susceptibility and resistance. Host abscesses are often devoid of oxygen. This is often the result of neutrophil recruitment and activity. We have determined that during anaerobic growth, the bacterial pathogen Staphylococcus aureus alters results in alters the production of S. aureus toxins and increases biofilm formation. Iron restriction also causes S. aureus to ferment. Lactoferrin prevents S. aureus for acquiring iron in the udder. Therefore, host environments that promote fermentative growth, such as the udder, could be causing S. aureus to alter the expression of it's virulence repertoire. OBJECTIVE 2: Characterization and manipulation of virulence factors of mastitis pathogens for enhancing host defenses. Biofilm formation is thought to be prerequisite for Staphylococcus aureus infections. Bacteria within biofilms can cause chronic and reoccurring infections. We have discovered that S. aureus responds to fermentative growth by increasing biofilm formation. The biofilm formation phenotype is the result of increased expression of fibronectin biding protein and cell lysis. Cell lysis resulted in DNA release, which contributed to intracellular adhesion. We identified two regulatory systems (SaeSR and SrrAB) that respond to decreased cellular respiration. Both regulatory systems are required for S. aureus pathogenesis. SaeSR is required for exotoxin production. Therefore, environments that promote fermentative metabolism increase the expression of virulence factors via altered SaeSR and SrrAB output. Staphylococcus aureus is a major cause of bovine mastitis--a disease that results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infectious disease related death in the USA. Therefore, the research conducted by my group impacts the health and pocketbooks of all Americans.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Mashruwala A.A., Boyd J.M., Investigating the role(s) of SufT and the domain of unknown function 59 (DUF59) in the maturation of iron-sulfur proteins. Current Genetics. 2017 Jun. pp 1-8. PMID: 28589301
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Mashruwala A.A., Gries C.M., Scherr T.D., Kielian T., Boyd J.M., SaeRS is responsive to the cellular respiratory status and regulates fermentative biofilm formation in Staphylococcus aureus. Infection and Immunity. nfect Immun. 2017 Jul 19;85(8). pii: e00157-17. doi: 10.1128/IAI.00157-17.
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Tanner A.W., Carabetta V.J., Martinie R.J., Mashruwala A.A., Boyd J.M., Krebs C., Dubnau D., The RicAFT (YmcA-YlbF-YaaT) complex carries two [4Fe-4S]2+ clusters and may respond to redox changes. Molecular Microbiology. 2017 Jun;104(5):837-850. 14 pages + supplemental material. PMID: 28295778
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Roberts C., Al-Tameemi H.M., Mashruwala A.A., Rosario-Cruz Z., Chauhan U., Sause W., Torres V.J., Boyd J.M., The Suf iron-sulfur cluster biosynthetic system is essential for Staphylococcus aureus viability and defective Fe-S cluster biosynthesis results in broad metabolic defects and decreased survival in neutrophils. Infection and Immunity. 2017 May 23;85(6). 19 pages. PMID: 28320837
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Mashruwala A.A., Van de Guchte A., Boyd J.M., Impaired respiration elicits SrrAB-dependent programmed cell lysis and biofilm formation in Staphylococcus aureus. eLife. 2017 Feb 21;6. 29 pages + supplemental material. PMID: 28221135
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Mashruwala A. A., Boyd J.M., The Staphylococcus aureus SrrAB regulatory system modulates hydrogen peroxide resistance factors, which imparts protection to aconitase during aerobic growth. PLoS One. 2017 Jan 18;12(1):e0170283. 30 pages + supplemental material. PMID: 28099473
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Mashruwala A.A., Roberts C., Bhatt S., May K.L., Carroll R.K., Shaw L.N., Boyd J.M., Staphylococcus aureus SufT: an essential iron-sulfur cluster assembly factor in cells experiencing a high-demand for lipoic acid. Molecular Microbiology. 2016 Dec;102(6):1099-1119. 21 pages + supplemental material. PMID: 27671355
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Choby J.E., Mike L.A., Mashruwala A.A., Dutter B.F., Dunman P.M., Sulikowski G.A., Boyd J.M.*, Skaar E.P.*, A small-molecule inhibitor of iron-sulfur cluster assembly uncovers a link between virulence regulation and metabolism in Staphylococcus aureus. Cell Chemical Biology. 2016 Nov 17;23(11):1351-1361. 11 pages + supplemental material. PMID:27773628
|
Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Staphylococcus aureus is a major cause of bovine mastitis--a disease that results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infectious disease related death in the USA. The research conducted by my group impacts the health and pocketbooks of all Americans. The findings from our research are of great interest to the public, scientists studying infection biology, research extension specialists attempting to prevent bovine mastitis,and physicians treating S. aureus infections. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The funds provided by this project have allowed me to purchase supplies for the training of three Ph.D. students, two M.S. students and two undergraduate students. These funds have also allowed my students to generate data that has resulted in a number of publications, which facilitated public awareness resulting requested speaking engagements and poster presentations. One student recently graduated with her Ph.D. and is now a full-time employee of an internationally recognized pharmaceutical company. the experiences that I have gained by training these students and the manuscripts that we authored have also greatly helped my professional development. How have the results been disseminated to communities of interest?We have published three peer-reviewed manuscripts in top journals this year and we have two more manuscripts in revision. Because these studies were partially funded by the NIH two of those papers are open access so the public can easily read about our findings. We paid for the third manuscript to be publically available through an open access journal. All of the papers directly related to the work that was funded by this NIFA project. The graduate and undergraduate students have presented their findings at 14 times at scientific meetings. I have also presented the results from the work funded by the NIFA project in two lectures to graduate students at Rutgers University. My group also participated in the annual Rutgers day celebration and used the platform to educate the public about the impact of microbiology on their daily lives. What do you plan to do during the next reporting period to accomplish the goals?This upcoming year we plan to focus our attention on determining how copper, an antimicrobial metal, prevents that growth of S. aureus. We will also focus our work to determine how S. aureus detoxifies this metal. A second goal is to better understand the mechanism(s) of anaerobic biofilm formation. Biofilms are a pre-requisite for S. aureus infections and biofilm-associated cells are considered the etiologic agents of recurrent Staphylococcus aureus infections. We will examine the effect of small molecule inhibitors of anaerobic biofilm formation.
Impacts
What was accomplished under these goals?
We spent most of last year examining how Staphylococcus aureus synthesizes small inorganic cofactors called iron-sulfur clusters. These protein-associated cofactors are sensitive to oxidative stress. We found that the process of building of FeS clusters is essential for S. aureus survival. Importantly, the machinery that S. aureus uses to build and maintain FeS clusters is different that that used by mammals. These data suggest that FeS cluster assembly is an important antibiotic target in this organism and other bacteria. We subsequently characterized one small molecule that inhibited the assembly of FeS clusters in proteins in S. aureus. This molecule was the first synthesized small molecule described to inhibit this process. These findings can and will be used by others to synthesize alternate molecular mimics of this small molecule with greater potency against S. aureus. We also characterized a protein that is present in nearly all organisms (SufT) that functions in the synthesis of FeS proteins. We found that this protein was necessary for the assembly of FeS proteins when the demand for FeS cluster synthesis is high. In total, our findings have uncovered a antibiotic target, characterized a small molecule that inhibitor of this target, and provided insight into the mechanisms by which S. aureus builds and maintains FeS cluster requiring proteins.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Mashruwala A.A., Bhatt S., Poudel S., Boyd E.S., and Boyd J.M. The DUF59 containing protein SufT is involved in the maturation of iron-sulfur (FeS) proteins during conditions of high FeS cofactor demand in Staphylococcus aureus. PLoS Genetics. 2016 Aug. PMID: 27517714
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Choby J.E., Mike L.A., Mashruwala A.A., Dutter B.F. Dunman, P.M., Sulikowski G.A., Boyd J.M. *, and Skaar E.P.* A small molecule inhibitor of iron-sulfur cluster assembly is toxic to Staphylococcus aureus in an Sae-dependent manner. Accepted. Cell Chemical Biology 2016 Nov. PMID:27773628
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Mashruwala A.A., Roberts C., Bhatt S. May K.L., Carroll R.K., Shaw L.N., Boyd J.M. Staphylococcus aureus SufT: an essential iron-sulfur cluster assembly factor in cells experiencing a high-demand for lipoic acid. Mol. Micro. 2016 Sep. PMID: 27671355
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Eveleigh D.E., H�ggblom M., and Boyd J.M. The early challenges of antibiotic discovery. Microbe. 2015 Nov; 10 (11): 449-450.
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Staphylococcus aureus is a major cause of bovine mastitis--a disease that results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infectious disease related death in the USA. The research conducted by my group impacts the health and pocketbooks of all Americans. The findings from our research are of great interest to the public, scientists studying infection biology, research extension specialists attempting to prevent bovine mastitis,and physicians treating S. aureus infections. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The funds provided by the USDA have provided money to purchase supplies, which aided in training the following students. 3 Ph.D. candidates 2 M.S. candidates 2 undergraduate researchers The funds provided have allowed my students to generate data that has resulted in a number of publications, which facilitated public awareness resulting requested speaking engagements. This has greatly helped my professional development and the professional development of the students. How have the results been disseminated to communities of interest?We have published four peer-reviewed papers in top journals this year. Because these studies were partially funded by the NIH two of those papers are open access so the public can easily read about our findings. All of the papers directly relate to the work that was funded by this NIFA project. I have presented our data in two international meetings and a handful of local meetings. The graduate and undergraduate students have presented their findings at numerous local meetings and one national meeting. I have also presented the results from the work funded by the NIFA project in two lectures to graduate students at Rutgers University, as well as, at Wake Forest University, the University of Kaiserslautern (Germany), Montana State University, Rutgers University, and the University of Delaware. My group also participated in the annual Rutgers day celebration and used the platform to educate the public about the impact of microbiology on their daily lives. What do you plan to do during the next reporting period to accomplish the goals?We will continue to examine small molecules that may alter the function of the two regulatory systems that we have discovered to be necessary for biofilm formation. We will also determine the mechanisms by which the regulatory systems alter biofilm formation and the dispersal of biofilms. We have found that DNA is a major component of the biofilm matrix, but how the DNA exits the cells is unknown. We will test the hypothesis that cell lysis is resulting in DNA and protein release providing matrix component to enhance biofilm formation. We will also continue to examine the role of copper ions in killing bacterial cells and the mechanisms by which S. aureus detoxifies copper atoms.
Impacts
What was accomplished under these goals?
Staphylococcus aureus is a major cause of bovine mastitis, which results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infections and is a major cause of infectious disease related death in the USA. Humans can acquire S. aureus infections from the infected cattle (zoonotic disease). These infections can be difficult to treat, in part, because many S. aureus strains are resistant to antibiotics commonly used in medical clinics. Infections caused by microbes that are resistant to antimicrobials result in prolonged hospital stays, increased financial burden, and increased mortality. Therefore, the results produced from the research conducted by this NIFA project impacts the health and pocketbooks of all Americans. Work in our lab strives to identify novel cellular targets to treat or prevent staphylococcal infections. Specifically, we examine the mechanisms of intracellular metal processing and the mechanisms by which S. aureus senses and responds to environmental perturbations. Work by our group has found that the disruption or modulation of these processes results in cell death or decreased virulence. These data confirm that these processes could be targeted by antimicrobial therapy. The overarching goal of our work is to identify unique and validated antimicrobial targets and develop antimicrobials that inhibit or alter the functions of these targets to prevent or cure infections. Objective 1: Characterization of host mechanisms associated with mastitis susceptibility and resistance. We have been actively examining the effect of copper (Cu) on S. aureus survival. Cu is used as a staphylococcal control agent and host defenses systems use Cu to prevent and clear S. aureus infections. We have identified two genes within the ACME pathogenesis island, which provide resistance to Cu. Importantly, this mobile DNA element is located adjacent to the mec antimicrobial resistance island and it is hypothesized the selection for the ACME pathogenesis island results in selection for the mec island, and therefore, multiple drug resistances. We have found that S. aureus strains lacking these Cu processing factors are much more susceptible to Cu intoxication. One of the proteins is a Cu binding protein and the other pumps Cu from the cytosol to the outside of the cell. These data provide information about how S. aureus circumvents host defense systems, which could result in mastitis formation. These data also suggest that Cu intoxication may be providing a selective pressure to increase antimicrobial resistance, which would be a paradigm shift in our thinking of how S. aureus acquired antimicrobial resistance encoded in mobile DNA. Objective 2: Characterization and manipulation of virulence factors of mastitis pathogens for enhancing host defenses. Biofilms are collections of cells that form a plaque rendering them more resistant to killing by the host and antimicrobial agents. Biofilms are thought to be a prerequisite for S. aureus infections. We have identified two regulatory systems that are required for biofilm formation. Work by others have found that both of these regulatory systems are also required for pathogenesis in animal models of infection. We have identified the stimuli for the regulatory systems and we have found that stimulating these regulatory systems results in programed cell death and increased biofilm formation. These studies have identified the molecular triggers for two regulatory systems that are vital for S. aureus pathogenesis. Continuation of these studies will allow us to determine the molecular mechanisms by which S. aureus forms and disperses biofilms, which are critical processes for infection. Objective 3. Assessment and application of new technologies that advance mastitis control, milk quality and dairy food safety. As mentioned above, we have identified regulatory systems that modulate biofilm formation and biofilm dispersal. We have also identified the molecular triggers that these regulatory systems respond to. We are currently attempting to modulate the behavior of the cells using small molecules that mimic the stimuli for these regulatory systems. If successful, these small molecules could be applied to modulate the behavior of S. aureus in the context of infection to disperse or prevent biofilms.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Rosario-Cruz Z., Chahal H.K., Mike L.A., Skaar E.P., Boyd J.M. Bacillithiol has a role in Fe-S cluster biogenesis in Staphylococcus aureus. Mol Microbiol. 2015 Oct;98(2):218-42. doi: 10.1111/mmi.13115. Epub 2015 Jul 30. PMID: 26135358
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Rosario-Cruz Z., Boyd J.M. Physiological roles of bacillithiol in intracellular metal processing. Curr Genet. 2016 Feb;62(1):59-65. doi: 10.1007/s00294-015-0511-0. Epub 2015 Aug 11.PMID 26259870
- Type:
Book Chapters
Status:
Published
Year Published:
2016
Citation:
Mashruwala A.A., Boyd J.M. De Novo assembly of plasmids using yeast recombinational cloning. Methods Mol Biol. 2016;1373:33-41. doi: 10.1007/7651_2015_275. PMID 26194707
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Gandhi S., Rosario-Cruz Z*., Boyd JM. Copper homeostasis in Staphylococcus aureus. University of Sao Paulo International Symposium of Scientific Initiation. San Paulo, Brazil. 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Earle C., Mashruwala, A.A., van de Gucht, A., Boyd J.M. Regulation of the Clp proteases by SrrAB in Staphylococcus aureus. University of Sao Paulo International Symposium of Scientific Initiation. San Paulo, Brazil. 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Rosario-Cruz Z*., Gandhi S., Boyd JM. Copper homeostasis in Staphylococcus aureus. Meeting of the New Jersey Antimicrobial Resistance Working Group. Piscataway, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A.A., van de Guchte*, A., Boyd J.M. Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. Meeting of the New Jersey Antimicrobial Resistance Working Group. Piscataway, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Paper. Mashruwala A.A.*, and Boyd J.M. Cellular respiration as a trigger for programmed cell death and biofilm formation in Staphylococcus aureus. New Jersey American Society of Microbiology Meeting in Miniature (Theobald Smith Society). New Brunswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A.A., van de Guchte*, A., Boyd J.M. Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. New Jersey American Society of Microbiology Meeting in Miniature (Theobald Smith Society). New Brunswick, NJ 2015
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Rosario-Cruz Z*., Gandhi S., Boyd JM. Copper homeostasis in Staphylococcus aureus. New Jersey American Society of Microbiology Meeting in Miniature (Theobald Smith Society). New Brunswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A., Earle,C.*, van de Guchte A., and Boyd JM. Regulation of Clp proteases by SrrAB in Staphylococcus aureus. Aresty Undergraduate Resarch symposium. Rutgers University. 2015
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Gandhi S*., Rosario-Cruz Z., Boyd JM. Copper homeostasis in Staphylococcus aureus. Aresty Undergraduate Research symposium. Rutgers University, New Brunswick, NJ 2015
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A., Earle,C.*, van de Guchte A., and Boyd JM. Regulation of Clp proteases by SrrAB in Staphylococcus aureus. New Jersey American Society of Microbiology Meeting in Miniature (Theobald Smith Society). New Brunswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A.A., van de Guchte*, A., Boyd J.M. Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. William Patterson Undergraduate Research Symposium. Wayne, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A.A*., van de Guchte, A., Boyd J.M. Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. Rutgers Microbiology Symposium. New Bruswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Mashruwala A.A., Jasim H*., Boyd J.M. TsrkR is required for thermal homeostatus in Staphylococcus aureus. Rutgers Microbiology Symposium. New Bruswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Zuelay Rosario-Cruz* and Boyd J.M. Copper homeostasis in Staphylococcus aureus. Rutgers Microbiology Symposium. New Bruswick, NJ 2015.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Paper. Boyd J.M., Investigating the role of SufT in Fe-S cluster biogenesis. Eastern Meeting on Fe-S cluster Biogenesis.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Paper. Boyd J.M., Targeting essential cellular processes and behavior modification. Rutgers antimicrobial resistance working group. New Brunswick, NJ
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Paper. Boyd J.M., Fe-S cluster assembly in the human bacterial pathogen Staphylococcus aureus: A viable target for antimicrobial therapy? International meeting on Fe-S cluster biogenesis and regulation. Bergamo, Italy
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster. Rosario-Cruz Z*., Gandhi S., Boyd JM. Copper homeostasis in Staphylococcus aureus. American Society of Microbiology National Meeting. New Orleans, LA 2015.
|
Progress 10/01/13 to 09/30/14
Outputs
Target Audience: Researchers and stakeholders in the dairy industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This research has provided training opportunities for: 1 postdoctoral researcher 4 Ph.D. candidates 3 undergraduate researchers How have the results been disseminated to communities of interest? We have published five peer-reviewed papers this year. Four of those papers directly relate to the work that was funded by this USDA-NIFA project. My students and myself have presented the data in three international meetings. I have also presented the results from the work funded by the USDA-NIFA project in two lectures to graduate students at Rutgers University. My group also anticipated in the NJAESAg Field Day and educated the public about the impact of microbiology on their daily lives. What do you plan to do during the next reporting period to accomplish the goals? We plan to continue to examine the roles of the regulatory system in sensing the absence of a terminal electron acceptor and determine how increased biofilm formation under fermentative growth conditions occurs. The formation of a biofilm involves many virulence factors. In the next year we plan on: 1. identifying the molecules that indirectly interacs with the regulatory system, resulting in stimulation and biofilm formation. 2. determine the mechanisms by which stimulation of this regulatory system results in biofilm formation. 3. identify factors that are regulated by the regulatory system that aid in biofilm formation.
Impacts
What was accomplished under these goals?
The concentration of oxygen available for use by invading microorganismsas a terminal electron acceptor greatly varies in diverse tissue types. During the last year, we have focused on elucidating the mechanisms by which anaerobic metabolism triggers biofilm formation.Biofilms are groups of microorganisms that stick to a surface or each other. We have discovered that culturing Staphylococcus aureus in the absence of oxygen results in a robust increase in the amount of biofilm formed. Biofilm formation is an important aspect of virulence, suggesting that factors invoved in biofilm formation are virulence factors. The increase in biofilm formation witnessed in the absence of oxygen is decreased if cells are co-cultured with the alternate terminal electron acceptorn nitrate. These data suggest that the absence of a terminal electron acceptor, and not the absence of oxygen, results in the phenotypes witnessed. We have discovered that the increased biofilm formation is the result of increased cell death, cell lysis and DNA release. We have also identified a regulatory system that indirectly senses the absence of a terminal electron acceptor and trigger for programed cell death.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2014
Citation:
Mashruwala A.A., Pang Y.Y., Rosario-Cruz Z., Chahal H.K., Benson M.A., Anzaldi-Mike L.L., Skaar E.P., Torres V.J., Nauseef W.M., Boyd J.M. Nfu facilitates that maturation of iron-sulfur proteins and participates in virulence in staphylococcus aureus. Mol. Micro. 2014. PMID: 25388433
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Boyd E.S., Thomas K.M., Dai Y., Boyd J.M.*, Outten F.W.* interplay between oxygen and Fe-S cluster biogenesis: Insights from the Suf pathway. Biochemistry. 2014. Sep 11. PMID: 25153801
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
White M.J., Boyd J.M., Horswill A.R., Nauseef W.M., Phosphatidylinositol-specific phospholipase C contributes to survival of Staphylococcus aureus USA300 in human blood and neutrophils. Infect. Immun. 2014. Jan 22. PMID: 24452683
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. Molecular Genetics of Bacteria and Phages meeting, Madison WI, 2014.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Cellular respiration as a trigger for multicellular behavior in Staphylococcus aureus. International meeting on Gram positive pathogens, Omaha NE 2014.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Poster Zuelay Rosario-Cruz*, Laura Anzaldi Mike, Eric Skaar, and Boyd J.M., Investigating the role of Bacillithiol in iron-sulfur cluster metabolism in Staphylococcus aureus. American Society of Microbiology General Meeting. Boston, MA 2014.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Paper Zuelay Rosario-Cruz*, Laura Anzaldi Mike, Eric Skaar, and Boyd J.M., In vivo evidence suggesting a role for bacillithiol in iron-sulfur cluster metabolism in Staphylococcus aureus. Rutgers Joint Molecular Biosciences Graduate Student annual symposium. New Brunswick, NJ. 2014.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Poster Zuelay Rosario-Cruz*, Laura Anzaldi Mike, Eric Skaar, and Boyd J.M., Investigating the role of Bacillithiol in iron-sulfur cluster metabolism in Staphylococcus aureus. New Jersey American Society of Microbiology Meeting in Miniature (Theobald Smith Society), New Brunswick, NJ 2014.
|
Progress 10/01/12 to 09/30/13
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The accomplishments listed above were completed by two graduate students in my lab and one postdoctoral fellow. How have the results been disseminated to communities of interest? In the last year we have disseminated our results at the following meetings: Gordon conference on staphylococcual diseases (two poster presentations) Southest meeting on Fe-S cluster biogenesis (invited talk) presentations where given at the University of Wisconsin, Rutgers University-New Brunswick, and Rutgers University-Newark (invited talks) International Conference on Iron-Sulfur Cluster Biogenesis and Regulation (invited talk) Theobold Smith meeting of the New Jersey chapter of the American Societe of Microbiology (3 posters) Northeastern microbiologists meeting (one poster) What do you plan to do during the next reporting period to accomplish the goals? Define the roleof two regulatory systems in Staphylococcus aureus biofilm formation.
Impacts
What was accomplished under these goals?
Staphylococcus aureus is a prominent pathogen that requires iron (Fe) for host colonization and infection. Although much is known about Fe acquisition, little is known about intracellular Fe metabolism. Once internalized, S. aureus mobilizes the Fe to build inorganic cofactors called Fe-S clusters.Our research findings are consistent with the hypothesis that the process of synthesizing Fe-S clusters is essential for S. aureus survival. We provide genetic and biochemical evidence showing that the Nfu protein is a Fe-S cluster trafficking protein. An nfu mutant strain was used to examine the effects of defective Fe-S cluster metabolism on staphylococcal physiology and pathogenesis. An S. aureus strain defective in Fe-S cluster metabolism displayed decreased cellular fitness, an increased intracellular non-chelated Fe pool, and increased endogenous reactive oxygen species. We conclude that a combination of the above factors results in increased Fenton chemistry in the cell, leading to DNA damage and cell death. The nfu strain was more sensitive to reactive oxygen and reactive nitrogen species and consistent with our physiology studies the nfu mutant strain was more susceptible to oxidative killing by human polymorphonuclear leukocytes. Finally, using a mouse model of infection we found that effective Fe-S cluster metabolism is necessary for tissue colonization. The results presented herein establish Fe-S cluster metabolism gene products as indispensable components of staphylococcal pathogenesis and crucial to cell survival, thus highlighting them as attractive antimicrobial targets.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2014
Citation:
White M.J., Boyd J.M., Horswill A.R., Nauseef W.M., Phosphatidylinositol-specific phospholipase C contributes to survival of Staphylococcus aureus USA300 in human blood and neutrophils. Infect. Immun. 2014. Accepted.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Walker J.N., Spaulding A., Salgado-Pab�n W., Schlievert P.M., Boyd J.M., Horswill A.R., The Staphylococcus aureus ArlRS two-component system is a novel regulator of agglutination and pathogenesis. PLoS Pathogens. PLoS Pathog. 2013 Dec;9 PMID: 24367264.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Pang Y.Y., Schwartz J., Boyd J.M., Horswill AR, William M. Nauseef W.M., Methionine Sulfoxide Reductases Protect against Oxidative Stress in Staphylococcus aureus Encountering Exogenous Oxidants and Human Neutrophils. J Innate Immun. 2013
|
Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: Outputs Activities. Our group has spent the last year conducting experiments to examine how the mammalian pathogen Staphylococcus aureus responds to changes in its environment and nutrient availability. During the past year, Professor Boyd mentored one postdoctoral researcher, five graduate students, and four undergraduate students in laboratory research. Prof. Boyd aided these students in conducting experiments and analyzing results. Prof. Boyd co-designed the curricula and taught one-half of the lectures for a graduate class in microbial physiology. Prof. Boyd designed the curricula and taught a class for undergraduates on microbial physiology. Events. Prof. Boyd attended the East Coast Fe-S enzyme meeting in Blacksburg VA the Gordon Conference on Iron-Sulfur Enzymes, the 60th anniversary of Selman Waksman Nobel Prize meeting, and the International Conference on Gram-Positive Pathogens. Professor Boyd presented his work at all four meetings. One student attended the International Meeting on Gram Positive Pathogens and presented his work. Prof. Boyd and 3 graduate students attended the 2011 Rutgers University Microbiology Symposium and all four individuals presented their work. During the past year this project has allowed me to enter active collaborations with Victor Torres, New York University; Alex Horswill, University of Iowa; William Nauseef, University of Iowa; Greg Sommerville, University of Nebraska; Antonio Perik, Philipps-Universitat Marburg, Germany; William Belden, Rutgers University; Alastair McEwan, the University of Queensland; Eric Skaar, Vanderbilt University; and Ann Stock, Rutgers University/UMDNJ. In collaboration with William Belden, I devised a way to clone any gene into any plasmid, using a yeast genetic system. We will be seek a patent for this technology. Dissemination. Participated in Ag field day and educated the public about the impact of microbiology on their daily lives. Organized voter education seminar series to inform the public about the three microbes that we have nominated to become the state microbe of NJ. I also aided in designing a website that allows interested parties to educate themselves about the importance of microbes, that we isolated at Rutgers University, on their daily lives. The website can be found here: http://dbm.rutgers.edu/stateMicrobe.php PARTICIPANTS: Jeff Boyd. Principal investigator Harsimranjit K. Chahal. Postdoctoral researcher Ameya Mashruwala. Ph.D. student Zuelay Rosario-Cruz. Ph.D. student Shiven Bhatt. M.S. student Shiming Tang, M.S. student Adriana van de Guchte, Undergraduate researcher Bhavana Narala, Undergraduate researcher Valerie T. Raziano, Undergraduate researcher Colaborators. Victor Torres, New York University; Alex Horswill, University of Iowa; William Nauseef, University of Iowa; Greg Sommerville, University of Nebraska; Antonio Perik, Philipps-Universitat Marburg, Germany. William Belden, Rutgers University; Alastair McEwan, the University of Queensland Ann Stock, Rutgers University/UMDNJ. Eric Skaar, Vanderbilt University. Training or professional development. The following individuals were trained in my lab during the past year. Ameya Mashruwala. Ph.D. student Zuelay Rosario-Cruz. Ph.D. student Shiven Bhatt. M.S. student Shiming Tang, M.S. student Adriana van de Guchte, Undergraduate researcher Bhavana Narala, Undergraduate researcher Valerie T. Raziano, Undergraduate researcher Jeffery Matthews, Undergraduate researcher Eric Huselid, Rotating Ph.D. student Ibrahim Alsawaf, Rotating Ph.D. student TARGET AUDIENCES: Target audiences. Staphylococcus aureus is a major cause of bovine mastitis; a disease that results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infectious disease related death in the USA. The research conducted by my group impacts the health and pocketbooks of all Americans. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
For successful colonization of host tissues, S. aureus must acquire and effectively and efficiently metabolize iron (Fe). Although much is known about how S. aureus acquires Fe from the host, little is known about what happens to the iron once internalized. Much of the Fe that S. aureus internalizes is used to build small inorganic cofactors called iron-sulfur ([Fe-S]) clusters. The innate immune system of mammals produces chemical oxidents such as hydrogen peroxide, hypochlorous acid (bleach), superoxide, and nitric oxide to kill invading bacteria, in part, by damaging [Fe-S] clusters, which are required for numerous metabolic processes. Once the clusters are damaged the iron is released and can freely react with hydrogen peroxide to produce hydroxyl radicals that result in cell death. We created a series of mutant S. aureus strains that are defective in [Fe-S] cluster metabolism and used these stains to test the hypothesis that an S. aureus strain that is defective intracellular Fe metabolism has altered virulence properties. We have focus our efforts on two genes nfu (SAUSA300_0839) and sufT (SAUSA300_0875). The mutants had lower activity of the Fe-S cluster dependent enzymes aconitase and isopropylmalate isomerase. These mutant strains had destabilized and increased intracellular unchelated Fe pools and were sensitive to the chemicals hydrogen peroxide, superoxide and nitric oxide. Biochemical analysis found that the nfu protein bound a Fe-S cluster, but the type of Fe-S cluster is currently unknown. We used classic and molecular genetics to examine the Fe-S cluster interactome. We found that potentially two pathways exist to build Fe-S clusters. The SufT protein interacted with the Fe-S cluster trafficking proteins consistant with the hypothesis that it is involved in the late stages of Fe-S cluster insertion into apo-proteins. We have also found that a S. aureus strain defective in producing the small molecule bacillithiol is defective in intracellular metal metabolism. The bacillithiol mutant was sensitive to copper, cobalt, and cadmium. It also had a growth defect with out exogenous added iron.
Publications
- Chahal H.K., Outten F.W., and Boyd J.M. 2012. Fe-S cluster biogenesis in Archaea and Bacteria. Metals in Cells. Wiley Publishing. Editors Valarie Culotta and Robert Scott.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Outputs/Activities. Our group has spent the last year conducting experiments to examine how the mammalian pathogen Staphylococcus aureus responds to changes in its environment and nutrient availability. During the past year, Professor Boyd mentored four graduate students and four undergraduate students that worked in the laboratory. Prof. Boyd aided these students in conducting experiments and analyzing results. Prof. Boyd co-designed the curricula and taught one-half of the lectures for a graduate class in microbial physiology. Prof. Boyd served on the admissions and the curriculum development and standards committees for the microbial biology graduate program. Events. Prof. Boyd attended the 6th International Conference on Fe‐S Protein Biogenesis and Regulation, the 55th Wind River Conference on Prokaryotic Biology, and the International Conference on Gram-Positive Pathogens. Prof. Boyd and 3 graduate students attended the 2011 Rutgers University Microbiology Symposium and all four individuals presented their work. Services. Co-instructor of a graduate class in microbial physiology. Products. I co-designed the curricula for a graduate class in microbial physiology. During the past year this project has allowed me to enter active collaborations with Victor Torres, New York University; Alex Horswill, University of Iowa; William Nauseef, University of Iowa; Greg Sommerville, University of Nebraska; Antonio Perik, Philipps-Universitat Marburg, Germany; William Belden, Rutgers University; and Ann Stock, Rutgers University/UMDNJ. In collaboration with William Belden, I devised a way to clone any gene into any plasmid, using a yeast genetic system. We will be seek a patent for this technology. Events. I participated in the following conferences during the previous year: 2011 Microbiology Symposium - Microbiology at Rutgers University - Cultivating Traditions, Current Strength and Future Frontiers. New Brunswick, NJ; Iron-sulfur cluster metabolism and Staphylococcus aureus virulence. 55th Wind River Conference on Prokaryotic Biology, Estes Park, CO, USA; Iron-sulfur cluster metabolism and Staphylococcus aureus virulence 6th International Conference on Fe‐S Protein Biogenesis and Regulation, Oxford, UK. Dissemination. Participated in Ag field day and educated the public about the impact of microbiology on their daily lives. I gave a presentation at Fairleigh Dickinson University on S. aureus antibiotic resistance and disease. Research was present at three international meetings and one regional meeting. PARTICIPANTS: Project participants Jeff Boyd. Principal investigator Ameya Mashruwala. Laboratory technician/Ph.D. student Zuelay Rosario-Cruz. Ph.D. student Alison Morel. Rotating Ph.D. student Shiven Bhatt. M.S. student Benjamin Nuta. Undergraduate researcher Adriana van de Guchte, Undergraduate researcher Bhavana Narala, Undergraduate researcher Valerie T. Raziano, Undergraduate researcher Colaborators. Victor Torres, New York University; Alex Horswill, University of Iowa; William Nauseef, University of Iowa; Greg Sommerville, University of Nebraska; Antonio Perik, Philipps-Universitat Marburg, Germany. William Belden, Rutgers University; Ann Stock, Rutgers University/UMDNJ. Training or professional development. The following individuals were trained in my lab during the past year. Ameya Mashruwala. Ph.D. student Zuelay Rosario-Cruz. Ph.D. student Alison Morel. Rotating Ph.D. student Hamidah Raduwan. Rotating Ph.D. student Shiven Bhatt. M.S. student Benjamin Nuta. Undergraduate researcher Adriana van de Guchte, Undergraduate researcher Bhavana Narala, Undergraduate researcher Valerie T. Raziano, Undergraduate researcher Jeffery Matthews, Undergraduate researcher TARGET AUDIENCES: Staphylococcus aureus is a major cause of bovine mastitis; a disease that results in decreased milk yield and costs the dairy industry billions of dollars each year. S. aureus infections are also the leading cause of human infectious disease related death in the USA. The research conducted by my group impacts the health and pocketbooks of all Americans. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
For successful colonization of host tissues, S.aureus must acquire and effectively and efficiently metabolize iron (Fe). Although much is known about how S. aureus acquires Fe from the host, little is known about what happens to the iron once internalized. Much of the Fe that S. aureus internalizes is used to build small inorganic cofactors called iron-sulfur ([Fe-S]) clusters. The innate immune system of mammals produces chemical oxidents such as hydrogen peroxide, hypochlorous acid (bleach), superoxide, and nitric oxide to kill invading bacteria, in part, by damaging [Fe-S] clusters, which are required for numerous metabolic processes. Once the clusters are damaged the iron is released and can freely react with hydrogen peroxide to produce hydroxyl radicals that result in cell death. We created a series of mutant S. aureus strains that are defective in [Fe-S] cluster metabolism and used these stains to test the hypothesis that an S. aureus strain that is defective intracellular Fe metabolism has altered virulence properties. The mutant strains had destabilized and increased intracellular unchelated Fe pools and were sensitive to the chemicals hydrogen peroxide, superoxide and nitric oxide. The mutant strains also increased the produced a virulence factors such as toxins and enzymes that S. aureus used to detoxify host derived factors. One of these strains had decreased survival in human neutrophils and was found defective in colonizing liver tissue in a mouse model of infection. Collectively, these studies highlight the importance of [Fe-S] cluster metabolism for S. aureus pathogenesis and provide a framework to further elucidate pathways of intracellular Fe metabolism in this pathogenic bacterium.
Publications
- Boyd, J.M., Endrizzi, J.A., Hamilton, T.L., Christopherson, M.R., Downs, D.M., and Peters, J.W., 2011. FAD binding by ApbE protein from Salmonella enterica: a new class of FAD binding proteins. J. Bacteriol. Feb;193(4):887-95.
- Boyd, J.M., Teoh, W.P., and Downs, D.M. 2011. Decreased transport suppresses the growth defect of an apbC mutant on tricarballylate. J. Bacteriol.. Epub Nov 13.
- Abstracts Boyd J.M. 2011. An enemy at the gates: Examining the Staphylococcal neutrophil interface. 2011 Microbiology Symposium - Microbiology at Rutgers University - Cultivating Traditions, Current Strength and Future Frontiers. New Brunswick, NJ
- Jeff M. Boyd, Ameya Mashruwala, Victor Torres, Yun Yun Pang, and William Nauseef. 2011. Iron-sulfur cluster metabolism and Staphylococcus aureus virulence. 55th Wind River Conference on Prokaryotic Biology, Estes Park, CO, USA.
- Jeff M. Boyd, Ameya Mashruwala, Victor Torres, Yun Yun Pang, and William Nauseef. 2011. Iron-sulfur cluster metabolism and Staphylococcus aureus virulence 6th International Conference on Fe‐S Protein Biogenesis and Regulation, Oxford, UK.
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