Staphylococcus aureus is a major human pathogen that causes significant morbidity and mortality in both hospital- and community-acquired infections. The appearance of multidrug-resistant strains has compounded this problem, galvanizing efforts aimed at identifying novel therapeutic targets. One promising area for the development of novel antimicrobials is S. aureus heme metabolism, as heme acquisition is vital to staphylococcal pathogenesis. S. aureus acquires heme from the most abundant heme source in the host, hemoglobin contained within circulating erythrocytes. The process of heme uptake and metabolism is performed by the iron regulated surface determinant system (Isd). This is an application for continuation of a project to dissect the biochemical steps involved in S. aureus heme acquisition. Studies conducted during the present funding period indicate that S. aureus has evolved to preferentially recognize human hemoglobin over hemoglobin from other animal species. Once internalized, the intracellular fate of heme is dependent on the iron status of the bacterium. More specifically, iron starved S. aureus utilize heme as an iron source while heme is segregated intact to the bacterial membrane during iron replete conditions. Based on these fundamental discoveries, new studies are proposed to understand the mechanism and function of heme acquisition in S. aureus. Our current working model is that IsdB functions as the hemoglobin receptor that permits heme-iron uptake into the bacterial cytoplasm. During conditions of iron starvation, heme is degraded to free iron and an unprecedented small molecule which we have named staphylobilin. The regulation of heme degradation occurs through transcriptional and post-transcriptional mechanisms allowing S. aureus to tailor its heme utilization machinery to respond to changes in iron and heme levels. Under iron replete conditions, heme is not degraded but is instead used intact as a cofactor of membrane hemoproteins. This proposal focuses on testing this model for heme acquisition and metabolism in a series of three integrated Specific Aims. We will utilize genetics, inorganic chemistry, biochemistry, and animal infection experiments to (i) define the mechanism by which S. aureus preferentially utilizes human hemoglobin, (ii) determine the fate and function of heme and staphylobilin within bacterial cells, and (iii) provide a mechanistic understanding of how S. aureus regulates the Isd system in response to changes in nutrient availability. Results from these studies will yield a molecular blueprint of the heme-iron acquisition machinery in S. aureus, and permit the rational design of small molecule inhibitors for therapeutic intervention against S. aureus infection.

Public Health Relevance

S. aureus is one of the most significant threats to the public health of the United States. This proposal will provide mechanistic insights into the contribution of Isd-mediated heme-iron acquisition to the pathogenesis of S. aureus infections. Considering that components of the Isd system are found in numerous Gram positive pathogens, results accrued from these studies will be applicable across a variety of infectious diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI069233-08
Application #
8451261
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Huntley, Clayton C
Project Start
2006-02-01
Project End
2016-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
8
Fiscal Year
2013
Total Cost
$366,284
Indirect Cost
$131,284
Name
Vanderbilt University Medical Center
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Choby, Jacob E; Buechi, Hanna B; Farrand, Allison J et al. (2018) Molecular Basis for the Evolution of Species-Specific Hemoglobin Capture by Staphylococcus aureus. MBio 9:
Lopez, Christopher A; Skaar, Eric P (2018) The Impact of Dietary Transition Metals on Host-Bacterial Interactions. Cell Host Microbe 23:737-748
Cassat, James E; Moore, Jessica L; Wilson, Kevin J et al. (2018) Integrated molecular imaging reveals tissue heterogeneity driving host-pathogen interactions. Sci Transl Med 10:
Lentz, Christian S; Sheldon, Jessica R; Crawford, Lisa A et al. (2018) Identification of a S. aureus virulence factor by activity-based protein profiling (ABPP). Nat Chem Biol 14:609-617
Grunenwald, Caroline M; Bennett, Monique R; Skaar, Eric P (2018) Nonconventional Therapeutics against Staphylococcus aureus. Microbiol Spectr 6:
Lojek, Lisa J; Farrand, Allison J; Weiss, Andy et al. (2018) Fur regulation of Staphylococcus aureus heme oxygenases is required for heme homeostasis. Int J Med Microbiol 308:582-589
Peng, Hui; Zhang, Yixiang; Palmer, Lauren D et al. (2017) Hydrogen Sulfide and Reactive Sulfur Species Impact Proteome S-Sulfhydration and Global Virulence Regulation in Staphylococcus aureus. ACS Infect Dis 3:744-755
Noto, Michael J; Burns, William J; Beavers, William N et al. (2017) Mechanisms of pyocyanin toxicity and genetic determinants of resistance in Staphylococcus aureus. J Bacteriol :
Surdel, Matthew C; Horvath Jr, Dennis J; Lojek, Lisa J et al. (2017) Antibacterial photosensitization through activation of coproporphyrinogen oxidase. Proc Natl Acad Sci U S A 114:E6652-E6659
Lojek, Lisa J; Farrand, Allison J; Wisecaver, Jennifer H et al. (2017) Chlamydomonas reinhardtii LFO1 Is an IsdG Family Heme Oxygenase. mSphere 2:

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