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TitleA Structure-Based Mechanism for DNA Entry into the Cohesin Ring.
Journal, issue, pagesMol Cell, Vol. 79, Issue 6, Page 917-933.e9, Year 2020
Publish dateSep 17, 2020
AuthorsTorahiko 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 /
PubMed AbstractDespite 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 ...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.
External linksMol Cell / PubMed:32755595 / PubMed Central
MethodsEM (single particle)
Resolution3.94 - 27.0 Å
Structure data

EMDB-10870:
Cohesin complex with loader gripping DNA
Method: EM (single particle) / Resolution: 27.0 Å

10930

6yuf

EMDB-10930, PDB-6yuf:
Cohesin complex with loader gripping DNA
Method: EM (single particle) / Resolution: 3.94 Å

Chemicals

ChemComp-BEF:
BERYLLIUM TRIFLUORIDE ION

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

Source
  • Schizosaccharomyces pombe (fission yeast)
  • schizosaccharomyces pombe (strain 972 / atcc 24843) (yeast)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / Chromosome segregation Sister chromatid cohesion SMC complexes Cohesin ABC-ATPase Mis4-Scc2-NIPBL

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