The National Institute of General Medical Sciences (NIGMS) awarded a $375,420 Project Grant (CFDA 93.859 - Biomedical Research and Research Training) to the University of Texas Medical Branch at Galveston (UTMB) to investigate the structural and functional mechanisms of translation quality control by canonical and non-canonical protein factors in bacteria. The project aims to elucidate how the ribosome, release factor 3 (RF3), initiation factors 1 and 2 (IF1, IF2), elongation factor 4 (EF-4),...
This Project Grant award of $256,995.00 from the National Institute of Allergy and Infectious Diseases (NIAID) under the Allergy and Infectious Diseases Research program (CFDA 93.855) supports research to develop a generalizable strategy for linking quantitative measures of bacterial pathogen traits to host cell mutations. The principal investigator at Tufts University School of Medicine will use optical sequencing techniques to identify host cell gene mutations that impact the growth and iron...
This Project Grant award of $1,359,316 from the National Institute of Allergy and Infectious Diseases (NIAID), under the Allergy and Infectious Diseases Research program (CFDA 93.855), supports research to investigate the regulation of mistranslation by the tuberculosis-causing bacterium Mycobacterium tuberculosis (MTB). The goal is to understand how MTB uses regulation of translation error rates, or "mistranslation", to survive stressors including antibiotics and the host environment....
This Project Grant award, funded by the National Institute of General Medical Sciences (NIGMS) under the Biomedical Research and Research Training program (CFDA 93.859), aims to provide significant insights into the molecular mechanism of ribosomal subunit joining in bacteria. The $403,423 award, with a performance period from August 20, 2024 to July 31, 2027, will support research at the University of Arkansas that investigates how antibiotics impact the biophysical parameters governing subunit...
This Project Grant award from the National Institute of General Medical Sciences (NIGMS), under the Biomedical Research and Research Training (CFDA 93.859) program, provides $658,165 to Triad National Security, LLC to conduct advanced research on the effects of ions on mRNA elements, ribosomes, and antibiotic-RNA interactions. The research aims to gain a better understanding of the fundamental physics underlying ribosome operation and mRNA regulation, with a focus on the bacterial ribosome and...
The National Institute of Allergy and Infectious Diseases (NIAID) awarded a $456,000 Project Grant (CFDA 93.855 - Allergy and Infectious Diseases Research) to Northwestern University to conduct research on how the CAS2 ribonuclease protein in Legionella pneumophila (the bacterium that causes Legionnaires' disease) non-canonically controls the pathogen's thermal tolerance and virulence. The research aims to determine if CAS2 acts directly by binding and cleaving/degrading a portion of the HSPC2...
This Project Grant award from the National Institute of General Medical Sciences (NIGMS), under the Biomedical Research and Research Training (CFDA 93.859) program, provides $316,968.00 to Emory University to study the regulation, inhibition, and quality control mechanisms of ribosomes. The key research activities focus on three main themes: 1) understanding translation fidelity events, including miscoding, quality control, and mRNA frameshifting; 2) investigating ribosome rescue when...
This federal Project Grant award from the National Institute of General Medical Sciences (NIGMS), under the Biomedical Research and Research Training (CFDA 93.859) program, is providing $372,880 to The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. to conduct research on the regulation of protein synthesis during cellular stress. The key objectives of this 5-year project are to: 1) understand how the formation of the essential translation initiation factor eIF2...
This National Science Foundation (NSF) Biological Sciences program grant to Yale University, totaling $940,000 over 3 years beginning March 1, 2024, aims to systematically identify features of human and viral messenger RNAs (mRNAs) that regulate protein production. The project will employ a massively parallel reporter assay to quantify the impact of RNA modifications, structured RNA elements, and cell-specific factors on mRNA translation. Insights gained will provide a comprehensive...
This $1,200,000 Project Grant award from the National Science Foundation's Biological Sciences program supports research at the University of Illinois to elucidate the role of peptidyl-tRNA hydrolase (PTH) enzymes in bacterial protein synthesis quality control mechanisms. The 4-year project aims to use ribosome profiling and structural biology approaches to identify where PTH rescues stalled ribosomes and understand the underlying mechanistic principles. The research findings are expected to...
CHARACTERIZING HOST TRANSLATION INHIBITION BY LEGIONELLA PNEUMOPHILA - PROJECT SUMMARY TRANSLATION IS A DYNAMIC AND ENERGETICALLY DEMANDING FUNDAMENTAL PROCESS REQUIRING NUMEROUS FACTORS THAT PROMOTE AND REGULATE PROTEIN SYNTHESIS. DYSREGULATION OF TRANSLATION OFTEN OCCURS IN MANY HUMAN DISEASES. FOR EXAMPLE, MANY PATHOGENS PROMOTE THEIR OWN REPLICATION BY TARGETING THESE TRANSLATION FACTORS, THE RIBOSOME, AND REGULATORY MECHANISMS TO UNDERMINE TRANSLATION IN THEIR HOST CELL. ONE SUCH BACTERIUM IS LEGIONELLA PNEUMOPHILA, THE CAUSATIVE AGENT OF THE SEVERE PNEUMONIA CALLED "LEGIONNAIRES' DISEASE". THE INCIDENCE OF LEGIONELLA INFECTIONS IN THE UNITED STATES HAS INCREASED IN RECENT YEARS, NECESSITATING A BETTER UNDERSTANDING OF HOW THIS BACTERIUM INTERACTS WITH ITS HOST CELL. LEGIONELLA TRANSLOCATES HUNDREDS OF TOXIC PROTEINS, TERMED EFFECTORS, THAT SUBVERT MANY BIOCHEMICAL PROCESSES WITHIN ITS HOST. NOTABLY, AT LEAST ELEVEN OF THESE EFFECTORS INHIBIT TRANSLATION. THESE EFFECTORS EXERT THEIR FUNCTION THROUGH A RANGE OF MECHANISMS, AND MOST APPEAR TO BE ENZYMES THAT MODIFY TRANSLATION FACTORS TO INACTIVATE THEM WHILE OTHERS MARK PROTEINS FOR DEGRADATION. DESPITE SEVERAL STUDIES DESCRIBING THE MECHANISMS OF SOME OF THESE TRANSLATION-INHIBITING EFFECTORS, IT IS UNCLEAR WHY LEGIONELLA EMPLOYS THEM. THIS IS DRIVEN BY THE FACT THAT THE MECHANISMS OF SOME EFFECTORS REMAIN ELUSIVE BUT ALSO BECAUSE THEIR SEEMINGLY REDUNDANT NATURE HAS MADE IT DIFFICULT TO ISOLATE THEIR PHYSIOLOGICAL RELEVANCE. THIS PROPOSAL SEEKS TO CLOSE THE GAPS IN KNOWLEDGE REGARDING THE TRANSLATION-INHIBITING EFFECTORS BY USING BOTH SPECIFIC AND BROAD APPROACHES. FIRST, I WILL DETERMINE THE MECHANISM OF ONE OF THESE EFFECTORS, SIDL. BASED ON STRONG PRELIMINARY DATA, I PROPOSE A SET OF EXPERIMENTS TO CHARACTERIZE ITS FUNCTION AND IDENTIFY ITS MOLECULAR TARGET. SECOND, I PROPOSE A COMPREHENSIVE TEMPORAL ANALYSIS OF TRANSLATION INHIBITION DURING THE LEGIONELLA INFECTION CYCLE. LEGIONELLA HAS A ~24-HOUR INFECTION CYCLE AND SOME STUDIES SUGGEST THAT THE TRANSLATION-INHIBITING EFFECTORS ARE TEMPORALLY REGULATED; HOWEVER, A CAREFUL CHARACTERIZATION OF WHEN EACH EFFECTOR WORKS PRECLUDES A DEEPER UNDERSTANDING OF THEM. TOGETHER, THESE TWO APPROACHES WILL ESTABLISH A FRAMEWORK TO UNDERSTAND THE PHYSIOLOGICAL RELEVANCE OF THE TRANSLATION-INHIBITING EFFECTORS OF LEGIONELLA. FURTHERMORE, BECAUSE THESE EFFECTORS TARGET HOST CELL FACTORS, THIS WORK CAN REVEAL NOVEL INSIGHTS INTO OUR OWN CELL BIOLOGY.