A Structure-Based Mechanism for DNA Entry into the Cohesin Ring
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 betwe...
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| Published in: | Molecular cell Vol. 79; no. 6; p. 917 |
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| Main Authors: | , , , , , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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17.09.2020
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| ISSN: | 1097-4164, 1097-4164 |
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| Abstract | 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. |
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| AbstractList | 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.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. 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. |
| Author | Sousa, Joana S Snijders, Ambrosius P Rappsilber, Juri Chen, Zhuo A O'Reilly, Nicola O'Reilly, Francis J Nans, Andrea Higashi, Torahiko L Eickhoff, Patrik Flynn, Helen R Costa, Alessandro Uhlmann, Frank Papageorgiou, George Locke, Julia |
| Author_xml | – sequence: 1 givenname: Torahiko L surname: Higashi fullname: Higashi, Torahiko L organization: Chromosome Segregation Laboratory, The Francis Crick Institute, London NW1 1AT, UK – sequence: 2 givenname: Patrik surname: Eickhoff fullname: Eickhoff, Patrik organization: Macromolecular Machines Laboratory, The Francis Crick Institute, London NW1 1AT, UK – sequence: 3 givenname: Joana S surname: Sousa fullname: Sousa, Joana S organization: Macromolecular Machines Laboratory, The Francis Crick Institute, London NW1 1AT, UK – sequence: 4 givenname: Julia surname: Locke fullname: Locke, Julia organization: Macromolecular Machines Laboratory, The Francis Crick Institute, London NW1 1AT, UK – sequence: 5 givenname: Andrea surname: Nans fullname: Nans, Andrea organization: Structural Biology STP, The Francis Crick Institute, London NW1 1AT, UK – sequence: 6 givenname: Helen R surname: Flynn fullname: Flynn, Helen R organization: Proteomics STP, The Francis Crick Institute, London NW1 1AT, UK – sequence: 7 givenname: Ambrosius P surname: Snijders fullname: Snijders, Ambrosius P organization: Proteomics STP, The Francis Crick Institute, London NW1 1AT, UK – sequence: 8 givenname: George surname: Papageorgiou fullname: Papageorgiou, George organization: Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK – sequence: 9 givenname: Nicola surname: O'Reilly fullname: O'Reilly, Nicola organization: Peptide Chemistry STP, The Francis Crick Institute, London NW1 1AT, UK – sequence: 10 givenname: Zhuo A surname: Chen fullname: Chen, Zhuo A organization: Bioanalytics Unit, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, Germany – sequence: 11 givenname: Francis J surname: O'Reilly fullname: O'Reilly, Francis J organization: Bioanalytics Unit, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, Germany – sequence: 12 givenname: Juri surname: Rappsilber fullname: Rappsilber, Juri organization: Bioanalytics Unit, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, Germany; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, UK – sequence: 13 givenname: Alessandro surname: Costa fullname: Costa, Alessandro email: alessandro.costa@crick.ac.uk organization: Macromolecular Machines Laboratory, The Francis Crick Institute, London NW1 1AT, UK. Electronic address: alessandro.costa@crick.ac.uk – sequence: 14 givenname: Frank surname: Uhlmann fullname: Uhlmann, Frank email: frank.uhlmann@crick.ac.uk organization: Chromosome Segregation Laboratory, The Francis Crick Institute, London NW1 1AT, UK. Electronic address: frank.uhlmann@crick.ac.uk |
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| Keywords | ABC-ATPase cohesin SMC complexes chromosome segregation sister chromatid cohesion S. pombe DNA-protein crosslink mass spectrometry Mis4/Scc2/NIPBL DNA loop extrusion cryo-electron microscopy |
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| SubjectTerms | Adenosine Triphosphatases - genetics Cell Cycle Proteins - genetics Cell Cycle Proteins - ultrastructure Chromatids - genetics Chromatids - ultrastructure Chromosomal Proteins, Non-Histone - genetics Chromosomal Proteins, Non-Histone - ultrastructure Chromosome Segregation - genetics Cohesins Cryoelectron Microscopy DNA - genetics DNA - ultrastructure Nucleic Acid Conformation Protein Conformation Saccharomyces cerevisiae - ultrastructure Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - ultrastructure Sister Chromatid Exchange - genetics |
| Title | A Structure-Based Mechanism for DNA Entry into the Cohesin Ring |
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