跳转至内容
Merck
  • Ubiquitin-specific peptidase 20 regulates Rad17 stability, checkpoint kinase 1 phosphorylation and DNA repair by homologous recombination.

Ubiquitin-specific peptidase 20 regulates Rad17 stability, checkpoint kinase 1 phosphorylation and DNA repair by homologous recombination.

The Journal of biological chemistry (2014-06-14)
Ilanchezhian Shanmugam, Mohammad Abbas, Farhan Ayoub, Susan Mirabal, Manal Bsaili, Erin K Caulder, David M Weinstock, Alan E Tomkinson, Robert Hromas, Monte Shaheen
摘要

Rad17 is a subunit of the Rad9-Hus1-Rad1 clamp loader complex, which is required for Chk1 activation after DNA damage. Rad17 has been shown to be regulated by the ubiquitin-proteasome system. We have identified a deubiquitylase, USP20 that is required for Rad17 protein stability in the steady-state and post DNA damage. We demonstrate that USP20 and Rad17 interact, and that this interaction is enhanced by UV exposure. We show that USP20 regulation of Rad17 is at the protein level in a proteasome-dependent manner. USP20 depletion results in poor activation of Chk1 protein by phosphorylation, consistent with Rad17 role in ATR-mediated phosphorylation of Chk1. Similar to other DNA repair proteins, USP20 is phosphorylated post DNA damage, and its depletion sensitizes cancer cells to damaging agents that form blocks ahead of the replication forks. Similar to Chk1 and Rad17, which enhance recombinational repair of collapsed replication forks, we demonstrate that USP20 depletion impairs DNA double strand break repair by homologous recombination. Together, our data establish a new function of USP20 in genome maintenance and DNA repair.

材料
货号
品牌
产品描述

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
环己酰亚胺,大包装, from microbial, ≥94% (TLC)
Sigma-Aldrich
L -还原型谷胱甘肽, suitable for cell culture, BioReagent, ≥98.0%, powder
Sigma-Aldrich
环己酰亚胺 溶液, Ready-Made Solution, microbial, 100 mg/mL in DMSO, Suitable for cell culture
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
HEPES缓冲溶液, 1 M in H2O
Sigma-Aldrich
乙二胺四乙酸 溶液, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
环己酰亚胺,大包装, ≥90% (HPLC)
Sigma-Aldrich
乙二胺四乙酸, anhydrous, crystalline, BioReagent, suitable for cell culture
SAFC
HEPES
Sigma-Aldrich
乙二胺四乙酸, 99.995% trace metals basis
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
环己酰亚胺,大包装, Biotechnology Performance Certified
SAFC
HEPES
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
乙二胺四乙酸, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
L -还原型谷胱甘肽, ≥98.0%
Sigma-Aldrich
DL-丙氨酸, ≥99% (HPLC)
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
Sigma-Aldrich
DL-半胱氨酸, technical grade
Sigma-Aldrich
乙二胺四乙酸, BioUltra, anhydrous, ≥99% (titration)
Sigma-Aldrich
乙二胺四乙酸 二钠盐 溶液, BioUltra, for molecular biology, pH 8.0, ~0.5 M in H2O
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
Sigma-Aldrich
DL-丙氨酸, ≥99%, FCC, FG
Sigma-Aldrich
乙二胺四乙酸, purified grade, ≥98.5%, powder
Supelco
谷胱甘肽, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L -还原型谷胱甘肽, BioXtra, ≥98.0%
Sigma-Aldrich
乙二胺四乙酸, ≥98.0% (KT)
Millipore
环己酰亚胺 溶液, 0.1%, suitable for microbiology