The long-term objective of this research is to understand how bacterial cells maintain metal ion homeostasis. Essential metal ions must be obtained from the environment, transported into the cell, and trafficked to the correct intracellular destinations. When metal ions are limiting, high affinity uptake mechanisms are induced. When in excess, the cell induces the synthesis of efflux, detoxication, or storage mechanisms. The ability to obtain sufficient metal ions from the host is crucial to bacterial pathogens, metal uptake systems are highly expressed in vivo, and surface-exposed transport proteins are often targets for the host immune response. Disorders of metal ion homeostasis are also a major cause of human disease including anemias, Menkes', and Wilson's diseases, and several other neurodegenerative diseases. The genetic responses to changes in metal ion availability are coordinated by metalloregulatory proteins. In the Gram-positive bacterium Bacillus subtilis metal-specific regulators control the uptake of Mn (MntR), Fe (Fur), and Zn (Zur), and others regulate efflux mechanisms induced when metal ions are in excess (CueR, CzrA, ArsR, YdeT). We will measure the metal ion content and quotas needed for growth for both wild-type and mutants altered in metal ion homeostasis. We will test the hypothesis that adsorption of metal ions to the cell wall plays a role in both uptake and storage of metals and we will characterize pathways for Fe and Zn uptake. We will test the hypotheses that MrgA and Dps function in Fe storage and detoxification and that Zn-containing ribosomal proteins provide a major intracellular Zn store that can be mobilized under Zn limitation. Genetic and biochemical approaches will be used to further our understanding of metal-sensing by MntR, Fur, Zur, CueR, and ArsR family members by site-directed mutagenesis of metal ligands, in vivo analyses of altered function mutants, and structure-determination methods.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM059323-08
Application #
7391793
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Chin, Jean
Project Start
2000-02-01
Project End
2009-03-31
Budget Start
2008-04-01
Budget End
2009-03-31
Support Year
8
Fiscal Year
2008
Total Cost
$260,377
Indirect Cost
Name
Cornell University
Department
Microbiology/Immun/Virology
Type
Schools of Earth Sciences/Natur
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Chandrangsu, Pete; Loi, Vu Van; Antelmann, Haike et al. (2018) The Role of Bacillithiol in Gram-Positive Firmicutes. Antioxid Redox Signal 28:445-462
Chandrangsu, Pete; Rensing, Christopher; Helmann, John D (2017) Metal homeostasis and resistance in bacteria. Nat Rev Microbiol 15:338-350
Pi, Hualiang; Helmann, John D (2017) Ferrous iron efflux systems in bacteria. Metallomics 9:840-851
Makthal, Nishanth; Nguyen, Kimberly; Do, Hackwon et al. (2017) A Critical Role of Zinc Importer AdcABC in Group A Streptococcus-Host Interactions During Infection and Its Implications for Vaccine Development. EBioMedicine 21:131-141
VanderWal, Arica R; Makthal, Nishanth; Pinochet-Barros, Azul et al. (2017) Iron Efflux by PmtA Is Critical for Oxidative Stress Resistance and Contributes Significantly to Group A Streptococcus Virulence. Infect Immun 85:
Huang, Xiaojuan; Shin, Jung-Ho; Pinochet-Barros, Azul et al. (2017) Bacillus subtilis MntR coordinates the transcriptional regulation of manganese uptake and efflux systems. Mol Microbiol 103:253-268
Shin, Jung-Ho; Helmann, John D (2016) Molecular logic of the Zur-regulated zinc deprivation response in Bacillus subtilis. Nat Commun 7:12612
Pi, Hualiang; Patel, Sarju J; Argüello, José M et al. (2016) The Listeria monocytogenes Fur-regulated virulence protein FrvA is an Fe(II) efflux P1B4 -type ATPase. Mol Microbiol 100:1066-79
Chandrangsu, Pete; Helmann, John D (2016) Intracellular Zn(II) Intoxication Leads to Dysregulation of the PerR Regulon Resulting in Heme Toxicity in Bacillus subtilis. PLoS Genet 12:e1006515
Ji, Chang-Jun; Kim, Jung-Hoon; Won, Young-Bin et al. (2015) Staphylococcus aureus PerR Is a Hypersensitive Hydrogen Peroxide Sensor using Iron-mediated Histidine Oxidation. J Biol Chem 290:20374-86

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