Cel Gene expression is tightly regulated to allow rapid responses to cellular stimuli. In eukaryotes, the 3´ polyA tail of mRNAs plays key roles in post-transcriptional control. The Cleavage and Polyadenylation Factor (CPF), Ccr4–Not and Pan2–Pan3 multiprotein complexes add or remove polyA tails to regulate mRNA stability and efficiency of translation. They control expression of genes in the inflammatory response, miRNA-targeted gene silencing and expression of maternal mRNAs in oocyte development. These processes are deregulated in disease, including cancer and neurological disorders.Although the proteins that add and remove polyA tails are known, their mechanisms are poorly understood. My lab recently established methods to reconstitute the polyA machinery. This led to new insights into the link between transcription and polyadenylation, new understanding of the molecular mechanisms of deadenylation, and details of RNA recruitment.In this proposal, my objective is to understand the molecular basis for polyadenylation and deadenylation of specific mRNAs. This is now possible because of our novel methodological and biological advances. We will determine high-resolution structures of the polyA machinery using electron cryo-microscopy (cryo-EM), reconstitute their biochemical activities in vitro and study their in vivo functional roles. We use this integrated approach to study intact multiprotein complexes, not individual subunits or domains. This involves considerable technical challenges and an investment in developing high quality purifications and new structural methods. I will determine how the four enzymatic activities of CPF are coupled, the mechanisms by which Ccr4–Not targets specific RNAs, and the molecular basis for RNA recognition by Pan2–Pan3. Together, this will provide new biological and technological insights, leading to understanding of fundamental processes in gene expression and the role of polyA tails in disease. Dziedzina nauki natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesphysical sciencesopticsmicroscopyelectron microscopymedical and health sciencesclinical medicineoncologynatural sciencesbiological sciencesgeneticsRNAmedical and health sciencesbasic medicineneurology Słowa kluczowe mRNA polyA tail gene expression Program(-y) H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme Temat(-y) ERC-2016-COG - ERC Consolidator Grant Zaproszenie do składania wniosków ERC-2016-COG Zobacz inne projekty w ramach tego zaproszenia System finansowania ERC-COG - Consolidator Grant Instytucja przyjmująca UNITED KINGDOM RESEARCH AND INNOVATION Wkład UE netto € 2 016 697,00 Adres POLARIS HOUSE NORTH STAR AVENUE SN2 1FL Swindon Zjednoczone Królestwo Zobacz na mapie Region South West (England) Gloucestershire, Wiltshire and Bristol/Bath area Swindon Rodzaj działalności Research Organisations Linki Kontakt z organizacją Opens in new window Strona internetowa Opens in new window Uczestnictwo w unijnych programach w zakresie badań i innowacji Opens in new window sieć współpracy HORIZON Opens in new window Koszt całkowity € 2 016 697,00 Beneficjenci (1) Sortuj alfabetycznie Sortuj według wkładu UE netto Rozwiń wszystko Zwiń wszystko UNITED KINGDOM RESEARCH AND INNOVATION Zjednoczone Królestwo Wkład UE netto € 2 016 697,00 Adres POLARIS HOUSE NORTH STAR AVENUE SN2 1FL Swindon Zobacz na mapie Region South West (England) Gloucestershire, Wiltshire and Bristol/Bath area Swindon Rodzaj działalności Research Organisations Linki Kontakt z organizacją Opens in new window Strona internetowa Opens in new window Uczestnictwo w unijnych programach w zakresie badań i innowacji Opens in new window sieć współpracy HORIZON Opens in new window Koszt całkowity € 2 016 697,00