Several pathogenic spore-forming bacteria, including the causative agent of anthrax Bacillus anthracis, are inhaled or ingested as spores and resume growth and toxin production in the host after germination. The spore protein coat, which encapsulates the spore, is an important factor in the infectious cycle, because of its effective protective function and its regulatory role in the germination process by controlling access of germinants to receptors located in the inner membrane of the spore. The spore coat is a morphologically complex structure composed of approximately 60 different sporulation proteins that assemble around the nascent spore. In this proposal, we will use an integrated systems biology approach to map protein interaction networks and identify key regulatory hubs that nucleate spore coat assembly- which may serve as markers for spore detection and/or as targets for control of spore germination in applications against bioterrorism. In addition to its relevance to public health issues, we envision that our project will illustrate the value of systems biology approaches for investigating the assembly of complex structures. We propose to use high throughput protein localization screens to characterize the protein interaction networks that govern spore coat assembly in the model organism Bacillus subtilis. Specifically, we will use a library of fluorescent protein fusions to all of the coat proteins in B. subtilis, to define the spatiotemporal hierarchy of deposition of spore coat proteins around the spore, to identify which morphogenetic proteins are required for the recruitment of all spore coat proteins and learn the transcriptional and post-translational mechanisms underlying the regulation of coat assembly. The nature of the interactions established between pairs of coat proteins will be demonstrated using a large scale yeast two hybrid assay and various focused biochemical approaches. Finally, we will use computational tools to integrate the data gathered from these experiments and obtain a comprehensive representation of the spore coat protein interaction network that will serve as a template for the study of other spore-forming bacteria, particularly B. anthracis. 1 Project narrative Our research will derive new hypotheses and mechanisms for the resistance properties of Bacillus spores and the mechanisms that control spore germination. We anticipate that our results will have an impact on the design of therapeutic approaches to detect and eliminate pathogenic spore-forming bacteria.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081571-05
Application #
8307821
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Gaillard, Shawn R
Project Start
2008-09-01
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2012
Total Cost
$299,041
Indirect Cost
$103,021
Name
New York University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
041968306
City
New York
State
NY
Country
United States
Zip Code
10012
Arrieta-Ortiz, Mario L; Hafemeister, Christoph; Bate, Ashley Rose et al. (2015) An experimentally supported model of the Bacillus subtilis global transcriptional regulatory network. Mol Syst Biol 11:839
Serrano, Mónica; Gao, JinXin; Bota, João et al. (2015) Dual-specificity anti-sigma factor reinforces control of cell-type specific gene expression in Bacillus subtilis. PLoS Genet 11:e1005104
Abe, Kimihiro; Kawano, Yuta; Iwamoto, Keito et al. (2014) Developmentally-regulated excision of the SPβ prophage reconstitutes a gene required for spore envelope maturation in Bacillus subtilis. PLoS Genet 10:e1004636
Bate, Ashley R; Bonneau, Richard; Eichenberger, Patrick (2014) Bacillus subtilis Systems Biology: Applications of -Omics Techniques to the Study of Endospore Formation. Microbiol Spectr 2:
McKenney, Peter T; Driks, Adam; Eichenberger, Patrick (2013) The Bacillus subtilis endospore: assembly and functions of the multilayered coat. Nat Rev Microbiol 11:33-44
Cozy, Loralyn M; Phillips, Andrew M; Calvo, Rebecca A et al. (2012) SlrA/SinR/SlrR inhibits motility gene expression upstream of a hypersensitive and hysteretic switch at the level of σ(D) in Bacillus subtilis. Mol Microbiol 83:1210-28
de Francesco, Melissa; Jacobs, Jake Z; Nunes, Filipa et al. (2012) Physical interaction between coat morphogenetic proteins SpoVID and CotE is necessary for spore encasement in Bacillus subtilis. J Bacteriol 194:4941-50
McKenney, Peter T; Eichenberger, Patrick (2012) Dynamics of spore coat morphogenesis in Bacillus subtilis. Mol Microbiol 83:245-60
Kacmarczyk, Thadeous; Waltman, Peter; Bate, Ashley et al. (2011) Comparative microbial modules resource: generation and visualization of multi-species biclusters. PLoS Comput Biol 7:e1002228
Waltman, Peter; Kacmarczyk, Thadeous; Bate, Ashley R et al. (2010) Multi-species integrative biclustering. Genome Biol 11:R96

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