A structure-based mechanism for DNA entry into the cohesin ring
journal contributionposted on 2020-10-09, 12:05 authored by Torahiko L Higashi, Patrik Eickhoff, Joana S Sousa, Julia Locke, Andrea Nans, Helen R Flynn, Ambrosius P Snijders, George Papageorgiou, Nicola O'Reilly, Zhuo A Chen, Francis J O’Reilly, Juri Rappsilber, Alessandro Costa, Frank Uhlmann
Despite key roles in sister chromatid cohesion and chromosome organization, the mechanism by which cohesin rings are loaded onto DNA is still unknown. Here we combine biochemical approaches and cryoelectron microscopy (cryo-EM) to visualize a cohesin loading intermediate in which DNA is locked between two gates that lead into the cohesin ring. Building on this structural framework, we design experiments to establish the order of events during cohesin loading. In an initial step, DNA traverses an N-terminal kleisin gate that is first opened upon ATP binding and then closed as the cohesin loader locks the DNA against the ATPase gate. ATP hydrolysis will lead to ATPase gate opening to complete DNA entry. Whether DNA loading is successful or results in loop extrusion might be dictated by a conserved kleisin N-terminal tail that guides the DNA through the kleisin gate. Our results establish the molecular basis for cohesin loading onto DNA.
Crick (Grant ID: 10198, Grant title: Uhlmann FC001198) Crick (Grant ID: 10065, Grant title: Costa FC001065) Crick (Grant ID: 10013, Grant title: O'Reilly FC001999) Crick (Grant ID: 10011, Grant title: Snijders FC001999) European Research Council (Grant ID: 670412 - ChromatidCohesion, Grant title: ERC 670412 - ChromatidCohesion) European Research Council (Grant ID: 820102 - CRYOREP, Grant title: ERC 820102 - CRYOREP)
ABC-ATPaseDNA loop extrusionDNA-protein crosslink mass spectrometryMis4/Scc2/NIPBLS. pombeSMC complexeschromosome segregationcohesincryo-electron microscopysister chromatid cohesionAdenosine TriphosphatasesCell Cycle ProteinsChromatidsChromosomal Proteins, Non-HistoneChromosome SegregationCryoelectron MicroscopyDNANucleic Acid ConformationProtein ConformationSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsSister Chromatid ExchangeUhlmann FC001198Costa FC001065PCPRTSBFERM-ackDevelopmental Biology06 Biological Sciences11 Medical and Health Sciences