The virulence of many human bacterial pathogens is dependent on transkingdom nanomachines, including type III secretion systems (T3SSs), which act to directly deliver tens of virulence proteins, often referred to as effectors, into the cytosol of mammalian cells. Many critical gaps exist in our understanding of how type III secreted (T3S) proteins, referred to as effectors, are defined and delivered to the T3S apparatus (T3SA). While each pathogen injects its own unique set of effectors into hosts, components of their machines share a high degree of similarity. For several decades, the dogma has been that the effector secretion is dependent on small acidic T3S chaperones that bind to their N-terminal regions. These chaperones control the hierarchy of secretion of proteins by mediating their recruitment to the sorting platform, a complex that cycles between the cytosol and membrane-embedded T3SA. Interestingly, cognate chaperones have not yet been identified for the majority of T3S effectors, including those from intensively studied Salmonella, Yersinia, Shigella and pathogenic Escherichia T3SSs. Here, we present data that support the existence of a noncanonical T3SS chaperone-independent (CI) pathway likely conserved across numerous phylogenetically distinct T3SS families. Here, using the Shigella flexneri T3SS as a model system, we propose to: 1. Determine how T3S chaperones are recruited to the sorting platform. Using the Protein Interaction 2. Dissect the molecular mechanisms by which CI effectors are recognized and delivered to the T3SA. 3. Investigate the existence of a co-translational ATPase-independent type III secretion pathway. Together the proposed studies shown not only advance our understanding regarding how T3S effectors are defined and delivered to the T3SA, but also result in the identification of targets for the development of novel antimicrobial agents that target the virulence of the large family of Gram-negative bacterial pathogens whose virulence is dependent on a functional T3SS.

Public Health Relevance

Many human bacterial pathogens utilized specialized nanomachines to deliver virulence proteins directly into host cells. The goal of our proposal is to characterize commonalities in the strategies by which these proteins are recruited to these machines. These studies will not only advance our understanding of these pathogens cause disease, but also likely to identify new targets for the development of novel antimicrobials to treat infections caused by these pathogens, particular as multidrug variants of several are starting to emerge.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
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Bacterial Pathogenesis Study Section (BACP)
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Mills, Melody
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Massachusetts General Hospital
United States
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