Bacterial endospores are agents of infectious disease, potential bioweapons, and causes of food spoilage and poisoning. The most notorious among these is Clostridium difficile-associated disease (CDAD) which continues to become more widespread as well are more severe. Elimination of spores from contaminated sites or materials is challenging due to the extreme resistance of these cell types to conventional antimicrobial treatments. Spores that have been triggered to germinate and lose dormancy are much more susceptible to antimicrobial treatments. Activation of a spore's native germination apparatus is a viable strategy for increasing the effectiveness of spore-killing methods. Dormant spores contain highly efficient machinery for sensing favorable changes in their environment and triggering germination. However, even under ideal laboratory germination conditions, up to 5% of a spore population will remain dormant. These """"""""superdormant"""""""" spores are resistant to decontamination and are a potential cause of infection at later dates. The proposed research will catalog and quantify the proteomes of the membrane fractions of dormant, germinating, and superdormant spores of the species Bacillus anthracis, Bacillus subtilis, C. difficile, and Clostridium perfringens. Proteome similarities and differences between species will contribute to our understanding of the mechanics of spore germination signaling and progression. Proteome differences between rapidly-germinating and superdormant spores will indicate potential causes of superdormancy. This data will be used to develop models for how the signaling apparatus involved in initiation of germination contributes to superdormancy. Results of the studies will guide further studies on spore germination and the development of more efficient methods of spore decontamination.

Public Health Relevance

Dormant bacterial spores are agents of infectious disease, potential bioweapons, and causes of food spoilage and poisoning. If spores can be caused to germinate, then they are much more susceptible to decontamination treatments. This research will reveal details of the natural germination apparatus, so that better methods for triggering germination can be devised.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI088298-02
Application #
8020041
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Ranallo, Ryan
Project Start
2010-03-01
Project End
2013-08-31
Budget Start
2011-03-01
Budget End
2013-08-31
Support Year
2
Fiscal Year
2011
Total Cost
$223,286
Indirect Cost
Name
Virginia Polytechnic Institute and State University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
003137015
City
Blacksburg
State
VA
Country
United States
Zip Code
24061
Tan, Irene S; Weiss, Cordelia A; Popham, David L et al. (2015) A Quality-Control Mechanism Removes Unfit Cells from a Population of Sporulating Bacteria. Dev Cell 34:682-93
Bernhards, Casey B; Chen, Yan; Toutkoushian, Hannah et al. (2015) HtrC is involved in proteolysis of YpeB during germination of Bacillus anthracis and Bacillus subtilis spores. J Bacteriol 197:326-36
Jorgenson, Matthew A; Chen, Yan; Yahashiri, Atsushi et al. (2014) The bacterial septal ring protein RlpA is a lytic transglycosylase that contributes to rod shape and daughter cell separation in Pseudomonas aeruginosa. Mol Microbiol 93:113-28
Ho, Theresa D; Williams, Kyle B; Chen, Yan et al. (2014) Clostridium difficile extracytoplasmic function ? factor ?V regulates lysozyme resistance and is necessary for pathogenesis in the hamster model of infection. Infect Immun 82:2345-55
Chen, Yan; Ray, W Keith; Helm, Richard F et al. (2014) Levels of germination proteins in Bacillus subtilis dormant, superdormant, and germinating spores. PLoS One 9:e95781
Liu, Hualan; McCord, Kristin D; Howarth, Jonathon et al. (2014) Hypermotility in Clostridium perfringens strain SM101 is due to spontaneous mutations in genes linked to cell division. J Bacteriol 196:2405-12
Melville, Stephen; Craig, Lisa (2013) Type IV pili in Gram-positive bacteria. Microbiol Mol Biol Rev 77:323-41
Liu, Hualan; Bouillaut, Laurent; Sonenshein, Abraham L et al. (2013) Use of a mariner-based transposon mutagenesis system to isolate Clostridium perfringens mutants deficient in gliding motility. J Bacteriol 195:629-36
Kumar, R Siva Sai; Hendrick, William; Correll, Jared B et al. (2013) Biochemistry and physiology of the ? class carbonic anhydrase (Cpb) from Clostridium perfringens strain 13. J Bacteriol 195:2262-9
Hartman, Andrea H; Liu, Hualan; Melville, Stephen B (2011) Construction and characterization of a lactose-inducible promoter system for controlled gene expression in Clostridium perfringens. Appl Environ Microbiol 77:471-8