Listeria monocytogenes is a Gram positive facultative intracellular bacterium that is capable of invading a wide range of host cell types and replicating in the cytosol. Proteins which mediate invasion, vacuolar escape, and survival in the host cytosol are all transcriptionally controlled by the positive regulatory factor PrfA. PrfA belongs t the cAMP receptor protein (Crp) family of transcription factors, which are activated upon binding small molecule cofactors. To date, a cofactor has not been identified for PrfA, although it is believed that a signal(s) specific to the host cell cytosol triggers PrfA activation, leading to virulence gene expression. We performed a forward genetic screen to identify factors that allow Lm to recognize its cytosolic localization and activate transcription of virulence genes. Our screen for 'cytosol sensing'was extremely successful, as we identified many genes that have severe defects in PrfA-dependent gene regulation, but have never before been associated with virulence. Two transposon mutants were chosen for further investigation, and each displayed a 3-5 log defect in colonization of the spleens and livers of intravenously infected mice. Experiments detailed in this application aim to dissect how these mutations regulate virulence gene expression in Listeria.

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Listeria is a model intracellular pathogen that specifically increases virulence gene expression upon entry into the host cytosol. The host-specific factor(s) that cue intracellular bacteria as to their cytosolic localization have not yet been identified. Experiments described in this application aim to identify the mechanisms by which intracellular pathogens sense the host cytosol environment and respond by inducing virulence gene expression.

National Institute of Health (NIH)
Postdoctoral Individual National Research Service Award (F32)
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Special Emphasis Panel (ZRG1)
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Mills, Melody
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University of California Berkeley
Schools of Arts and Sciences
United States
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Reniere, Michelle L; Whiteley, Aaron T; Hamilton, Keri L et al. (2015) Glutathione activates virulence gene expression of an intracellular pathogen. Nature 517:170-3
Pensinger, Daniel A; Aliota, Matthew T; Schaenzer, Adam J et al. (2014) Selective pharmacologic inhibition of a PASTA kinase increases Listeria monocytogenes susceptibility to β-lactam antibiotics. Antimicrob Agents Chemother 58:4486-94