Department of Molecular and Cell Biology and Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA 94720

How intracellular pathogens sense their intracellular environment to activate virulence gene expression

Intracellular pathogens collectively cause an enormous amount of world-wide morbidity and mortality.  In my lab, we study Listeria monocytogenes as a very amenable intracellular pathogen as a model for both bacterial pathogenesis and host response.  L. monocytogenes is a Gram-positive, food-borne bacterium that lives a biphasic lifestyle, cycling between a saprophytic existence in the environment and as an intracellular pathogen of mammals. In today’s lecture, I will ask how L. monocytogenes recognizes and responds to the intracellular environment.  I will begin by describing a novel genetic strategy to identify bacterial mutants that fail to up-regulate a virulence factor (ActA) that is not expressed in the environmental phase of growth but becomes the most highly expressed bacterial protein during intracellular growth.  Most of the mutants identified in this screen were in genes that control redox regulation including a gene that encodes glutathione synthase (gshF).  It turns out that both bacterially and to a lesser degree, host-derived glutathione, is the co-factor that activates the central transcriptional virulence activator, PrfA.  In culture, we could recapitulate this response by the addition of glutathione and surprisingly, by adding one of many different reducing agents to the growth media.  The addition of reducing agents caused the up-regulation of gshF, but we still did not know what the precise biological cue was to activate gshF expression.  GshF mutants were 200-fold less virulent in mice while a mutation in PrfA that is locked in its fully active configuration (referred to as PrfA*) completely rescued virulence of a gshF mutant. This led us to look for additional mutants that formed small plaques in tissue culture, but were rescued by the introduction of a PrfA* mutation.  Among the genes identified in this screen was gloA, which encodes glyoxalase A, a component of a glutathione-dependent methylglyoxal (MG) detoxification system. MG is a toxic byproduct of metabolism, which if accumulated, causes DNA damage and protein glycation. As a facultative intracellular pathogen, L. monocytogenes must protect itself from MG produced by its own metabolic processes and that of its host. The gloA mutants grew normally in broth but were sensitive to MG and severely attenuated upon IV infection in mice, but were fully rescued in a PrfA* background. We demonstrate that transcriptional activation of gshF increased upon MG challenge in vitro, yet gloA mutants had decreased levels of GSH, presumably because GSH reacted irreversibly with MG. These data suggest that MG is a host cue that leads to GshF production and activation of PrfA. 

Location: Zoom Virtual Meeting, details to be emailed to Branch Members

For non-members, watch the meeting on our YouTube Channel

780th Monthly Meeting
September 21, 2020
100th Year Celebration of the Eastern PA Branch ASM
Speakers: James Poupard, Alan Evangelista, Irving Nachamkin, Sunny Shin, Michele Kutzler

781st Monthly Meeting
October 26, 2020
Joe Zackular, University of Pennsylvania
“Microbial cooperation enhances Clostridioides difficile pathogenesis”

782nd Monthly Meeting
November 23rd, 2020
Daniel Portnoy, UC Berkeley
“How intracellular pathogens sense their intracellular environment to activate virulence gene expression”

December 4th 2020
Infection and Immunity Forum

783rd Monthly Meeting
January 25th, 2021
John Mekalanos, Harvard Medical School
“Protein covariance networks reveal interactions important to the emergence of SARS coronaviruses as human pathogens“

784th Monthly Meeting
February 22nd, 2021
Isabell Coppens, Johns Hopkins
“Catching What You Eat: The Secret of the Intracellular Tenure of Toxoplasma gondii

785th Monthly Meeting
March 22nd , 2021
Invited speaker

786th Monthly Meeting
April 26th, 2021
Invited speaker

787th Monthly Meeting
May 24th, 2021
Student Chapter Sponsored Meeting
Student speakers at 4-5pm, Invited Speaker 5-6pm

788th Monthly Meeting
June 14th, 2021
Norman Willett Memorial Lecture
Amy Vollmer, Swarthmore College
“Microbiology: Nevertheless, We Persisted”
(Sponsored by the Education Committee)

Joseph Zackular, Ph.D.
Assistant Professor, Department of Pathology and Laboratory Medicine
University of Pennsylvania Perelman School of Medicine
Children’s Hospital of Philadelphia

Microbial cooperation enhances Clostridioides difficile pathogenesis

Clostridioides difficile is the most commonly reported nosocomial pathogen in the United States and an urgent public health threat worldwide. The primary risk factor for C. difficile infection of the gastrointestinal tract is antibiotic use, which reduces colonization resistance to C. difficile by perturbing the resident gut microbiota. Despite the well-established link between the gut microbiota and susceptibility to C. difficile infection, the impact of polymicrobial interactions on C. difficile pathogenesis is not well understood.  In this talk, we will explore the effect of microbial cooperation on C. difficile virulence and investigate the molecular mechanisms of cross talk between C. difficile and the gut microbiota.

Congratulations to the organizers, speakers and participants for a successful program celebrating our 100th anniversary of the Eastern PA Branch ASM.

Introduction to the Meeting by Branch President Dieter M. Schifferli, Ph.D., Dr.Med.Vet., DVM and ASM President Vic DiRita, Ph.D., and talk by branch member and archivist James Poupard PhD

History Antimicrobial Resistance by branch member Alan Evangelista Ph.D., and History of Diagnostics by branch member Irving Nachamkin DrPH, MPH

Legionnaires’ Disease by branch member Sunny Shin Ph.D. and History of Vaccine Development by branch member Michele Kutzler Ph.D.

Question and Answer Session and Closing with moderators Toby Eisenstein Ph.D. and Linda Miller, Ph.D.