Purified Smc5/6 Complex Exhibits DNA Substrate Recognition and Compaction
| Autor: | Clara Aicart-Ramos, Fernando Moreno-Herrero, Pilar Gutiérrez-Escribano, Holger B. Kramer, Juri Rappsilber, Alex Montoya, Roger Solé-Soler, Kyle L. Morris, Silvia Hormeño, Jordi Torres-Rosell, Francis J. O’Reilly, Luis Aragón, Julene Madariaga-Marcos, Ricardo Aramayo |
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| Přispěvatelé: | Wellcome Trust, Generalitat de Catalunya, Agència de Gestió d'Ajuts Universitaris i de Recerca, London Institute of Medical Sciences, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Medical Research Council (UK), European Research Council, European Commission, Comunidad de Madrid, Ministerio de Economía y Competitividad (España) |
| Rok vydání: | 2020 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Smc5/6 purification DNA substrate recognition Chromosomal Proteins Non-Histone Condensin Mitosis Cell Cycle Proteins Saccharomyces cerevisiae Smc5/6 holocomplex Article Biophysical Phenomena 03 medical and health sciences chemistry.chemical_compound electron mmicroscopy 0302 clinical medicine ATP hydrolysis Electron microscopy Humans Interphase Molecular Biology 030304 developmental biology Adenosine Triphosphatases Magnetic tweezers 0303 health sciences DNA compaction Cohesin biology Sumoylation Single-molecule Cell Biology Electron mmicroscopy Ubiquitin ligase DNA-Binding Proteins chemistry Multiprotein Complexes Eukaryotic chromosome fine structure Biophysics biology.protein Plectoneme stabilisation 030217 neurology & neurosurgery DNA |
| Zdroj: | Molecular Cell Recercat. Dipósit de la Recerca de Catalunya instname Recercat: Dipósit de la Recerca de Catalunya Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya) Gutierrez-Escribano, P, Hormeño, S, Madariaga-Marcos, J, Solé-Soler, R, O’Reilly, F J, Morris, K, Aicart-Ramos, C, Aramayo, R, Montoya, A, Kramer, H, Rappsilber, J, Torres-Rosell, J, Moreno-Herrero, F & Aragon, L 2020, ' Purified Smc5/6 complex exhibits DNA substrate recognition and compaction ', Molecular Cell, vol. 80, no. 6, pp. 1039-1054.e6 . https://doi.org/10.1016/j.molcel.2020.11.012 Digital.CSIC. Repositorio Institucional del CSIC Repositorio Abierto de la UdL Universitad de Lleida |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2020.11.012 |
| Popis: | Summary 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. Graphical Abstract Highlights • Purification of enzymatically active Smc5/6 yeast holocomplex • Smc5/6 coiled-coils exhibit a folded conformation • Smc5/6 stabilizes DNA plectonemes • Smc5/6 compacts DNA against low forces in an ATP-dependent manner Gutierrez-Escribano et al. purify the entire Smc5/6 holocomplex, retaining full enzymatic function. The Smc5/6 complex compacts DNA against low force and stabilizes DNA crosses present in supercoiled and catenated DNA. These findings indicate that many Smc5/6 functions occur through stabilization of DNA tertiary structure. |
| Databáze: | OpenAIRE |
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