Regulation of VSV mRNA translation

About This Grant

Project Summary Project Description: Viruses are obligate intracellular parasites, as they require access to cellular translation machinery for viral protein synthesis. The order Mononegavirales contains non-segmented negative sense RNA viruses (nsNSV). These viruses transcribe positive sense, capped and polyadenylated mRNA that are indistinguishable from host transcripts. Due to this, it is assumed that these viruses use canonical translation machinery. However, the mRNA of these viruses possesses characteristics, such as extremely short 5’UTRs and strong stem-loop structures, that would preclude canonical translation initiation and scanning methods. Using a series of auxin-inducible degron (AID) systems, we have systematically probed the requirements of individual translation factors for protein synthesis. Vesicular stomatitis virus (VSV) is a member of the Mononegavirales order. Its translation has been the most studied of the nsNSVs. We have demonstrated that VSV mRNAs translate independently of factors necessary for canonical translation, leading us to believe that viral translation occurs through an alternative mechanism. This proposal aims to characterize the mechanism of VSV translation, a first step to expanding the knowledge of nsNSV translation. Long-term, this work will contribute to increasing the basic understanding of viral replication as well as identifying targets for the development of antivirals. Training Plan: I will receive guidance and feedback from my sponsor, Dr. Shawn Lyons, and co-sponsor, Dr. John Connor. In the lab, I will develop and refine my skills in biochemistry, translational research methods, and virology. Participation in regular lab meetings and journal clubs will hone my scientific and critical thinking skills, while attendance at departmental seminars will broaden my understanding of biochemistry and cell biology. Furthermore, I will gain expertise in RNA biology through monthly RNA club meetings, providing a specialized area of focus. I will regularly exercise my presentation skills, presenting in various fostering effective scientific communication skills. This comprehensive training will equip me with a strong foundation in biochemistry, virology, and RNA biology, positioning me for success in my current research endeavor. Further, this will provide me with the expertise necessary to obtain a postdoctoral training fellowship, which will guide me toward my ultimate goal of becoming a principal investigator in an academic setting. Environment: The Lyons lab is well-equipped with ample bench space, with various machines at our disposal. These include a Chemidoc and TurboBlot Transfer system, for western blotting, and a BioComp, for polysome analysis. We have access to shared spaces that accomadate cold storage and tissue culture. Within the building, we have access to several microscopes and a BioRAD NGC chromatography system, for protein purification. Additional equipment is available through various core facilities on Boston University’s Medical School and Charles River campuses.