PXD024160 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding |
Description | Eukaryotic cells employ three SMC 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. |
HostingRepository | PRIDE |
AnnounceDate | 2023-05-14 |
AnnouncementXML | Submission_2023-05-14_14:12:02.829.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | BarbaraSteigenberger |
SpeciesList | scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; |
ModificationList | No PTMs are included in the dataset |
Instrument | Q Exactive HF |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2021-02-12 06:06:54 | ID requested | |
1 | 2021-07-02 04:12:22 | announced | |
⏵ 2 | 2023-05-14 14:12:03 | announced | 2023-05-14: Updated project metadata. |
Publication List
Dataset with its publication pending |
Keyword List
ProteomeXchange project tag: EPIC-XS |
submitter keyword: ATPase, condensin, loop extrusion, chromosome segregation,XL-MS, Smc5, SMC, Smc6, genome maintenance, cohesin |
Contact List
R. A.Scheltema |
contact affiliation | Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Centre, Utrecht, The Netherlands. |
contact email | R.A.Scheltema@uu.nl |
lab head | |
BarbaraSteigenberger |
contact affiliation | MPI of Biochemistry |
contact email | steigenberger@biochem.mpg.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD024160
- Label: PRIDE project
- Name: Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding