Dataset Information

SPOP mass spec in different treatment

ABSTRACT: 293T cells were transfected with S, Flag, and Biotin-binding-protein-(streptavidin)-binding-peptide (SFB) triple-tagged backbone vector or SFB-tagged SPOP. Twenty-four h after transfection, control cells were treated with DMSO (Group 1) and SFB-SPOP transfected cells were treated with DMSO (Group 2) or DNA damaging agents including mitomycin C (MMC, 1 µM) (Group 3), camptothecin (CPT, 50 nM) (Group 4), cisplatin (10 µM) (Group 5) and etoposide (10 µM) (Group 6) for 24 h. The cells were lysed by NETN buffer (20 mM Tris-HCl, pH8.0, 100 mM NaCl, 1mM EDTA, 0.5% Nonidet P-40) with 50 mM β-glycerophosphate, 10 mM NaF, and 1μg/mL pepstatin-A at 4°C for 3 h, followed by tandem affinity purification using streptavidin beads and S tag beads and LC-MS. A total of six groups of cell lysate were analyzed (n=6), including one group of cells transfected with empty vector and treated with DMSO as a control.

ORGANISM(S): Homo Sapiens

SUBMITTER: Jian Ma

PROVIDER: PXD028210 | iProX | Mon Sep 06 00:00:00 BST 2021

REPOSITORIES: iProX

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Publications

SPOP mutation induces replication over-firing by impairing Geminin ubiquitination and triggers replication catastrophe upon ATR inhibition.

Ma Jian J Shi Qing Q Cui Gaofeng G Sheng Haoyue H Botuyan Maria Victoria MV Zhou Yingke Y Yan Yuqian Y He Yundong Y Wang Liguo L Wang Yuzhuo Y Mer Georges G Ye Dingwei D Wang Chenji C Huang Haojie H

Nature communications 20211001 1

Geminin and its binding partner Cdt1 are essential for the regulation of DNA replication. Here we show that the CULLIN3 E3 ubiquitin ligase adaptor protein SPOP binds Geminin at endogenous level and regulates DNA replication. SPOP promotes K27-linked non-degradative poly-ubiquitination of Geminin at lysine residues 100 and 127. This poly-ubiquitination of Geminin prevents DNA replication over-firing by indirectly blocking the association of Cdt1 with the MCM protein complex, an interaction requi ...[more]

PMID: 34599168

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Dataset Information

SPOP mass spec in different treatment

Publications

SPOP mutation induces replication over-firing by impairing Geminin ubiquitination and triggers replication catastrophe upon ATR inhibition.

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