Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast.
Cell Cycle Proteins
/ metabolism
Centromere
/ metabolism
Chromosomal Instability
Chromosomal Proteins, Non-Histone
/ metabolism
Chromosome Segregation
DNA-Binding Proteins
/ metabolism
F-Box Proteins
/ metabolism
Protein Domains
Proteolysis
SKP Cullin F-Box Protein Ligases
/ metabolism
Saccharomyces cerevisiae
/ genetics
Saccharomyces cerevisiae Proteins
/ metabolism
Ubiquitin-Protein Ligase Complexes
/ metabolism
Ubiquitin-Protein Ligases
/ metabolism
Ubiquitination
Journal
PLoS genetics
ISSN: 1553-7404
Titre abrégé: PLoS Genet
Pays: United States
ID NLM: 101239074
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
10
06
2019
accepted:
03
01
2020
revised:
20
02
2020
pubmed:
8
2
2020
medline:
12
5
2020
entrez:
8
2
2020
Statut:
epublish
Résumé
Restricting the localization of the histone H3 variant CENP-A (Cse4 in yeast, CID in flies) to centromeres is essential for faithful chromosome segregation. Mislocalization of CENP-A leads to chromosomal instability (CIN) in yeast, fly and human cells. Overexpression and mislocalization of CENP-A has been observed in many cancers and this correlates with increased invasiveness and poor prognosis. Yet genes that regulate CENP-A levels and localization under physiological conditions have not been defined. In this study we used a genome-wide genetic screen to identify essential genes required for Cse4 homeostasis to prevent its mislocalization for chromosomal stability. We show that two Skp, Cullin, F-box (SCF) ubiquitin ligases with the evolutionarily conserved F-box proteins Met30 and Cdc4 interact and cooperatively regulate proteolysis of endogenous Cse4 and prevent its mislocalization for faithful chromosome segregation under physiological conditions. The interaction of Met30 with Cdc4 is independent of the D domain, which is essential for their homodimerization and ubiquitination of other substrates. The requirement for both Cdc4 and Met30 for ubiquitination is specifc for Cse4; and a common substrate for Cdc4 and Met30 has not previously been described. Met30 is necessary for the interaction between Cdc4 and Cse4, and defects in this interaction lead to stabilization and mislocalization of Cse4, which in turn contributes to CIN. We provide the first direct link between Cse4 mislocalization to defects in kinetochore structure and show that SCF-mediated proteolysis of Cse4 is a major mechanism that prevents stable maintenance of Cse4 at non-centromeric regions, thus ensuring faithful chromosome segregation. In summary, we have identified essential pathways that regulate cellular levels of endogenous Cse4 and shown that proteolysis of Cse4 by SCF-Met30/Cdc4 prevents mislocalization and CIN in unperturbed cells.
Identifiants
pubmed: 32032354
doi: 10.1371/journal.pgen.1008597
pii: PGENETICS-D-19-00928
pmc: PMC7032732
doi:
Substances chimiques
CDC4 protein, S cerevisiae
0
CSE4 protein, S cerevisiae
0
Cell Cycle Proteins
0
Chromosomal Proteins, Non-Histone
0
DNA-Binding Proteins
0
F-Box Proteins
0
MET30 protein, S cerevisiae
0
Saccharomyces cerevisiae Proteins
0
Ubiquitin-Protein Ligase Complexes
EC 2.3.2.23
SKP Cullin F-Box Protein Ligases
EC 2.3.2.27
Ubiquitin-Protein Ligases
EC 2.3.2.27
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1008597Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM066164
Pays : United States
Organisme : NHGRI NIH HHS
ID : R01 HG005084
Pays : United States
Organisme : NHGRI NIH HHS
ID : R01 HG005853
Pays : United States
Organisme : CIHR
ID : FDN-143264
Pays : Canada
Déclaration de conflit d'intérêts
NO authors have competing interests.
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