NSF Award
2106417
With this award, the Chemistry of Life Processes Program in the Division of Chemistry, Professor Sarah Michel of the University of Maryland Baltimore will investigate the roles of native and exogenous metals on zinc-mediated RNA regulation. Zinc is a critical metal found in living systems and plays an important role in RNA regulation. This role includes associating with proteins called Zinc Fingers (ZF). Under certain biological conditions (e.g. inflammation) and certain environmental conditions (e.g. pollution), other metals are present at elevated levels in the cellular environment and affect RNA regulation. The PI will study how such exogenous metals affect the function of ZFs in RNA regulation. Fundamental knowledge regarding the biochemical mechanisms of metal mediated RNA regulation by ZFs will be obtained. Graduate and undergraduate student participants will be trained in cutting-edge research approaches to determine the principles of metal co-factored protein regulation. The PI will continue SIMSI, a partnership with Notre Dame of Baltimore, that connects female graduate students with undergraduate women for career panels, mini-laboratory experiences, summer internships, and a ‘Minding your Metals’ activity. ‘Minding Your Metals’ was developed by the PI for students to test the water in Baltimore City as ‘citizen scientists.’ The PI will work with additional partners from UMB CURE Scholars (URM students from Baltimore City middle schools) and Bryn Mawr School (high school girls in Baltimore City) to expand ‘Minding Your Metals’ to survey the soil in Baltimore City and to make connections with Baltimore City’s industrial past. Graduate students will participate in the outreach efforts, which will be opportunities for them to develop science communication and teaching skills.<br/><br/>This proposal is focused on three zinc finger (ZF) proteins that belong to the CCCH class of ZFs, namely Tristetraprolin (TTP), cleavage and polyadenylation specificity factor 30 (CPSF30), and YTH1 ll. TTP contains two CCCH domains and regulates inflammation by controlling the levels of cytokines. TTP activity is extremely sensitive to the local cellular environment, including the presence of exogenous metals. A major goal of this work is to determine how exogenous metals drive TTP function during inflammation using molecular, cellular and proteomics approaches. These approaches include native ESI-MS, ICP-MS, XAS and NMR spectroscopies, fluorescence anisotropy, and IP/ICP-MS. CPSF30 contains five CCCH domains and one CCHC domain, and it regulates pre-mRNA processing. Dr. Michel has discovered that CPSF30 contains a 2Fe-2S domain and binds two unique RNA sequences. The research will focus on the identification of the specific domain that “houses” the Fe-S cluster and elucidation of its function as well as on answering the question how CPSF30 binds two distinct RNA target sequences. YTH1 is a closely related yeast homolog of CPSF30, which lacks the CCHC domain. Research on YTH1 will focus on the questions whether YTH1 contains a 2Fe-2S cluster in addition to zinc sites and identification of the protein’s RNA target(s). Another goal of this work is to determine the mechanisms of metal-mediated RNA recognition by CPSF30 and YTH1 and to answer questions regarding the evolution of this family of proteins and the commonality of the 2Fe-2S cluster. EPR and CD spectroscopies, HDX and MCO/MS, and fluorescence anisotropy will be used to answer these questions.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
