Spores are major mechanism by which the important nosocomial pathogen Clostridium difficile transmits disease, yet little is known about how this obligate anaerobe forms the resistant spore form. While preventing spore assembly would break the destructive cycle of infection and re-infection that characterizes C. difficile infections, no therapies currently target this developmental process. A major obstacle to developing such therapies is the absence of a basic understanding of how C. difficile forms a spore. Our long-term goal is to understand how C. difficile assembles the protective proteinaceous spore coat, which is necessary for functional spore formation. We recently identified the first spore morphogenetic proteins in the Clostridia as SpoIVA and SipL and showed that they directly interact. In this application, we aim to elucidate the molecular mechanisms by which SpoIVA and SipL coordinately regulate spore formation. We will use complementary genetic, biochemical, and cytological methods to determine the functional significance of SpoIVA-SipL complex formation during spore assembly, identify new SpoIVA and/or SipL interacting partners that regulate spore assembly, and localize SpoIVA and SipL during spore formation. By interrogating the mechanism by which SpoIVA and SipL interact and recruit coat proteins to the forespore during sporulation, the proposed studies will elucidate the functions of key regulators of spore formation, identify new morphogenetic proteins in the Clostridia, and develop new imaging methods for localizing proteins in live cells under anaerobic conditions. Since SipL is found exclusively in the Clostridia, these studies will expand our limited understanding of how spore assembly is regulated in the Clostridia. Completion of the proposed work will provide the mechanistic insight necessary for rationally designing screens that can identify C. difficile sporulation inhibitors and may also identify new potential therapeutic targets. As a result, these studies may lead to novel therapeutics that can prevent disease transmission by C. difficile and other clostridial spore-forming pathogens.

Public Health Relevance

Clostridium difficile is a major nosocomial pathogen that costs the US health care system >$1 billion to treat each year. Since the spore form of C. difficile mediates disease transmission and recurrence, understanding the molecular mechanisms by which the spore coat assembles to produce a viable spore has the potential to lead to therapeutics that can prevent C. difficile infection and spread.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI122232-03
Application #
9186980
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Ranallo, Ryan
Project Start
2016-07-14
Project End
2020-11-30
Budget Start
2016-12-01
Budget End
2017-11-30
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Tufts University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02111
Ribis, John W; Ravichandran, Priyanka; Putnam, Emily E et al. (2017) The Conserved Spore Coat Protein SpoVM Is Largely Dispensable in Clostridium difficile Spore Formation. mSphere 2:
Shen, Aimee; Fimlaid, Kelly A; Pishdadian, Keyan (2016) Inducing and Quantifying Clostridium difficile Spore Formation. Methods Mol Biol 1476:129-42
Fimlaid, Kelly A; Jensen, Owen; Donnelly, M Lauren et al. (2015) Regulation of Clostridium difficile Spore Formation by the SpoIIQ and SpoIIIA Proteins. PLoS Genet 11:e1005562
Shen, Aimee (2015) A Gut Odyssey: The Impact of the Microbiota on Clostridium difficile Spore Formation and Germination. PLoS Pathog 11:e1005157