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Purified Smc5/6 Complex Exhibits DNA Substrate Recognition and Compaction.
Gutierrez-Escribano, Pilar; Hormeño, Silvia; Madariaga-Marcos, Julene; Solé-Soler, Roger; O'Reilly, Francis J; Morris, Kyle; Aicart-Ramos, Clara; Aramayo, Ricardo; Montoya, Alex; Kramer, Holger; Rappsilber, Juri; Torres-Rosell, Jordi; Moreno-Herrero, Fernando; Aragon, Luis.
Afiliação
  • Gutierrez-Escribano P; Cell Cycle Group, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK.
  • Hormeño S; Department of Macromolecular Structures, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
  • Madariaga-Marcos J; Department of Macromolecular Structures, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
  • Solé-Soler R; Institut de Recerca Biomèdica de Lleida (IRBLLEIDA), Department of Ciències Mèdiques Bàsiques, Universitat de Lleida, Lleida, Spain.
  • O'Reilly FJ; Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, Germany; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, UK.
  • Morris K; Microscopy Facility, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK.
  • Aicart-Ramos C; Department of Macromolecular Structures, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
  • Aramayo R; Microscopy Facility, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK.
  • Montoya A; Biological Mass Spectrometry and Proteomics Facility, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK.
  • Kramer H; Biological Mass Spectrometry and Proteomics Facility, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK.
  • Rappsilber J; Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355 Berlin, Germany; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, UK.
  • Torres-Rosell J; Institut de Recerca Biomèdica de Lleida (IRBLLEIDA), Department of Ciències Mèdiques Bàsiques, Universitat de Lleida, Lleida, Spain.
  • Moreno-Herrero F; Department of Macromolecular Structures, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain. Electronic address: fernando.moreno@cnb.csic.es.
  • Aragon L; Cell Cycle Group, MRC London Institute of Medical Sciences (LMS), Du Cane Road, London W12 0NN, UK. Electronic address: luis.aragon@lms.mrc.ac.uk.
Mol Cell ; 80(6): 1039-1054.e6, 2020 12 17.
Article em En | MEDLINE | ID: mdl-33301732
Eukaryotic SMC complexes, cohesin, condensin, and Smc5/6, use ATP hydrolysis to power a plethora of functions requiring organization and restructuring of eukaryotic chromosomes in interphase and during mitosis. The Smc5/6 mechanism of action and its activity on DNA are largely unknown. Here we purified the budding yeast Smc5/6 holocomplex and characterized its core biochemical and biophysical activities. Purified Smc5/6 exhibits DNA-dependent ATP hydrolysis and SUMO E3 ligase activity. We show that Smc5/6 binds DNA topologically with affinity for supercoiled and catenated DNA templates. Employing single-molecule assays to analyze the functional and dynamic characteristics of Smc5/6 bound to DNA, we show that Smc5/6 locks DNA plectonemes and can compact DNA in an ATP-dependent manner. These results demonstrate that the Smc5/6 complex recognizes DNA tertiary structures involving juxtaposed helices and might modulate DNA topology by plectoneme stabilization and local compaction.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Saccharomyces cerevisiae / Proteínas de Ciclo Celular / Proteínas de Saccharomyces cerevisiae / Complexos Multiproteicos Limite: Humans Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Saccharomyces cerevisiae / Proteínas de Ciclo Celular / Proteínas de Saccharomyces cerevisiae / Complexos Multiproteicos Limite: Humans Idioma: En Ano de publicação: 2020 Tipo de documento: Article