Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding.

Details

Serval ID
serval:BIB_43B3495591E9
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding.
Journal
The EMBO journal
Author(s)
Taschner M., Basquin J., Steigenberger B., Schäfer I.B., Soh Y.M., Basquin C., Lorentzen E., Räschle M., Scheltema R.A., Gruber S.
ISSN
1460-2075 (Electronic)
ISSN-L
0261-4189
Publication state
Published
Issued date
02/08/2021
Peer-reviewed
Oui
Volume
40
Number
15
Pages
e107807
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
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.
Keywords
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, Protein Conformation, Saccharomyces cerevisiae/metabolism, Saccharomyces cerevisiae Proteins/chemistry, Saccharomyces cerevisiae Proteins/genetics, Saccharomyces cerevisiae Proteins/metabolism, Smc5/6, chromosome segregation, cohesion, condensin, loop extrusion
Pubmed
Web of science
Open Access
Yes
Create date
06/07/2021 11:08
Last modification date
24/02/2022 7:34
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