PXD029573-1

PXD029573 is an original dataset announced via ProteomeXchange.

Title	Role of Nse1 subunit of SMC5/6 complex as a ubiquitin ligase
Description	Structural Maintenance of Chromosomes (SMC) complexes - cohesin, condensin and SMC5/6 - are important for many aspects of the chromosomal organization. These complexes are composed of Smc and non-Smc element (Nse) subunits. The Nse1 subunit of SMC5/6 contains a variant RING domain, characteristic of ubiquitin ligases. Human NSE1 has been shown to possess ubiquitin ligase activity in vitro; however, other studies were unable to show such activity. Here, we confirm Nse1 ubiquitin-ligase activity in vitro using purified Schizosaccharomyces pombe proteins. We demonstrate that the Nse1 ligase activity is stimulated by Nse3 and Nse4, it specifically utilizes Ubc13/Mms2 E2 enzyme and Nse1 interacts directly with ubiquitin. We identify Nse1 mutation (R188E) that specifically disrupts its E3 activity, and show that the Nse1-dependent ubiquitination is particularly important under replication stress. Moreover, we identify Nse4 (lysine K181) as the first known SMC5/6-associated Nse1 substrate. Interestingly, abolition of Nse4 modification at K181 leads to suppression of DNA-damage sensitivity of other SMC5/6 mutants. Altogether, this study brings new evidence for Nse1 ubiquitin ligase activity, significantly advancing our under-standing of this enigmatic SMC5/6 function.
HostingRepository	PRIDE
AnnounceDate	2022-02-19
AnnouncementXML	Submission_2022-02-19_11:22:22.247.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Potesil
SpeciesList	scientific name: Schizosaccharomyces pombe 927; NCBI TaxID: 1264690; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	ubiquitination signature dipeptidyl lysine
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2021-11-04 10:17:38	ID requested
⏵ 1	2022-02-19 11:22:22	announced
2	2023-11-14 08:54:12	announced	2023-11-14: Updated project metadata.

10.3390/CELLS11010165;

submitter keyword: SMC5/6

Nse1, ubiquitin ligase, ubiquitination, liquid chromatography, mass spectrometry

Zbynek Zdrahal
contact affiliation	CEITEC—Central European Institute of Technology, Masaryk University, Brno, Czech Republic
contact email	zdrahal@sci.muni.cz
lab head
David Potesil
contact affiliation	Proteomics Core Facility and Research Group Proteomics CEITEC-MU Masaryk University Brno, Czech Republic
contact email	david.potesil@ceitec.muni.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD029573
     1. Label: PRIDE project
     2. Name: Role of Nse1 subunit of SMC5/6 complex as a ubiquitin ligase