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Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding
- Source :
- The EMBO Journal, Taschner, M, Basquin, J, Steigenberger, B, Schäfer, I B, Soh, Y M, Basquin, C, Lorentzen, E, Räschle, M, Scheltema, R A & Gruber, S 2021, ' Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding ', EMBO Journal, vol. 40, no. 15, e107807 . https://doi.org/10.15252/embj.2021107807, EMBO Journal, 40(15), 1. Nature Publishing Group, The EMBO journal, vol. 40, no. 15, pp. e107807
- Publication Year :
- 2021
-
Abstract
- Eukaryotic cells employ three SMC (structural maintenance of chromosomes) complexes to control DNA folding and topology. The Smc5/6 complex plays roles in DNA repair and in preventing the accumulation of deleterious DNA junctions. To elucidate how specific features of Smc5/6 govern these functions, we reconstituted the yeast holo‐complex. We found that the Nse5/6 sub‐complex strongly inhibited the Smc5/6 ATPase by preventing productive ATP binding. This inhibition was relieved by plasmid DNA binding but not by short linear DNA, while opposing effects were observed without Nse5/6. We uncovered two binding sites for Nse5/6 on Smc5/6, based on an Nse5/6 crystal structure and cross‐linking mass spectrometry data. One binding site is located at the Smc5/6 arms and one at the heads, the latter likely exerting inhibitory effects on ATP hydrolysis. Cysteine cross‐linking demonstrated that the interaction with Nse5/6 anchored the ATPase domains in a non‐productive state, which was destabilized by ATP and DNA. Under similar conditions, the Nse4/3/1 module detached from the ATPase. Altogether, we show how DNA substrate selection is modulated by direct inhibition of the Smc5/6 ATPase by Nse5/6.<br />Biochemical reconstitution and structural analyses provide insight into the architecture and substrate‐associated conformational changes of the yeast holo‐SMC5/6 complex.
- Subjects :
- Saccharomyces cerevisiae Proteins
DNA repair
Chromosomal Proteins, Non-Histone
Protein Conformation
Adenosine Triphosphate/metabolism
Cell Cycle Proteins/chemistry
Cell Cycle Proteins/genetics
Cell Cycle Proteins/metabolism
Chromosomal Proteins, Non-Histone/chemistry
Chromosomal Proteins, Non-Histone/genetics
Chromosomal Proteins, Non-Histone/metabolism
Cryoelectron Microscopy
Crystallography, X-Ray
DNA, Fungal/metabolism
Hydrolysis
Multiprotein Complexes/chemistry
Multiprotein Complexes/metabolism
Saccharomyces cerevisiae/metabolism
Saccharomyces cerevisiae Proteins/chemistry
Saccharomyces cerevisiae Proteins/genetics
Saccharomyces cerevisiae Proteins/metabolism
Smc5/6
chromosome segregation
cohesion
condensin
loop extrusion
ATPase
Neuroscience(all)
DNA Folding
Cell Cycle Proteins
Saccharomyces cerevisiae
Biology
Biochemistry
General Biochemistry, Genetics and Molecular Biology
Article
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Adenosine Triphosphate
ATP hydrolysis
Structural Biology
Immunology and Microbiology(all)
Smc5-Smc6 complex
Binding site
DNA, Fungal
Molecular Biology
030304 developmental biology
0303 health sciences
General Immunology and Microbiology
Biochemistry, Genetics and Molecular Biology(all)
General Neuroscience
DNA Replication, Repair & Recombination
Articles
chemistry
Multiprotein Complexes
biology.protein
Biophysics
030217 neurology & neurosurgery
DNA
Cysteine
Genetics and Molecular Biology(all)
Subjects
Details
- Language :
- English
- ISSN :
- 02614189
- Volume :
- 40
- Issue :
- 15
- Database :
- OpenAIRE
- Journal :
- EMBO Journal
- Accession number :
- edsair.doi.dedup.....5c17986ccfdfc0da9d0095f7c5afdede