Recruitment of trimeric proliferating cell nuclear antigen by G1-phase cyclin-dependent kinases following DNA damage with platinum-based antitumour agents.
Recruitment of trimeric proliferating cell nuclear antigen by G1-phase cyclin-dependent kinases following DNA damage with platinum-based antitumour agents.
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铂类抗肿瘤药物损伤 DNA 后，G1 期细胞周期蛋白依赖性激酶募集三聚体增殖细胞核抗原。
DOI：
10.1038/bjc.2013.613
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发表时间：
2013-10-29
影响因子：
8.8
通讯作者：
Siddik, Z. H.
中科院分区：
文献类型：
--
作者：
He, G.;Kuang, J.;Koomen, J.;Kobayashi, R.;Khokhar, A. R.;Siddik, Z. H.
在循环肿瘤细胞中，二元细胞周期蛋白依赖性激酶Cdk 4/细胞周期蛋白D或Cdk 2/细胞周期蛋白E复合物在DNA损伤后被p21抑制以诱导G1细胞周期停滞。然而，目前尚不清楚其他蛋白质是否也被招募到Cdk复合物中，或者它们的作用，并对此进行了研究。卵巢A2780肿瘤细胞暴露于铂基抗肿瘤剂1 R，2 R-二氨基环己烷（反式-二乙酰氨基）（二氯）铂（IV）（DAP），其以p21依赖性方式优先诱导G1期阻滞。通过凝胶过滤色谱、免疫印迹和质谱分析Cdk复合物。在对照肿瘤细胞中Cdk 4和Cdk 2复合物的活性形式具有约140 kDa的分子大小，当被DAP在G1检查点激活后抑制时，其增加至约290 kDa。蛋白质组学分析确定了Cdk，细胞周期蛋白，p21和增殖细胞核抗原（PCNA）的抑制复合物，和生物化学研究提供了明确的证据表明，在150 kDa的抑制复合物的增加是一致的p21依赖性的招聘PCNA作为一个三聚体，可能绑定到三个分子的p21。虽然p21单独足以抑制Cdk复合物，但PCNA对稳定p21至关重要。受p21抑制的G1 Cdk复合物也募集PCNA，其抑制复合物内p21的降解，从而抑制复合物内p21的活性。
In cycling tumour cells, the binary cyclin-dependent kinase Cdk4/cyclin D or Cdk2/cyclin E complex is inhibited by p21 following DNA damage to induce G1 cell-cycle arrest. However, it is not known whether other proteins are also recruited within Cdk complexes, or their role, and this was investigated. Ovarian A2780 tumour cells were exposed to the platinum-based antitumour agent 1R,2R-diaminocyclohexane(trans-diacetato)(dichloro)platinum(IV) (DAP), which preferentially induces G1 arrest in a p21-dependent manner. The Cdk complexes were analysed by gel filtration chromatography, immunoblot and mass spectrometry. The active forms of Cdk4 and Cdk2 complexes in control tumour cells have a molecular size of ∼140 kDa, which increased to ∼290 kDa when inhibited following G1 checkpoint activation by DAP. Proteomic analysis identified Cdk, cyclin, p21 and proliferating cell nuclear antigen (PCNA) in the inhibited complex, and biochemical studies provided unequivocal evidence that the increase in ∼150 kDa of the inhibited complex is consistent with p21-dependent recruitment of PCNA as a trimer, likely bound to three molecules of p21. Although p21 alone was sufficient to inhibit the Cdk complex, PCNA was critical for stabilising p21. G1 Cdk complexes inhibited by p21 also recruit PCNA, which inhibits degradation and, thereby, prolongs activity of p21 within the complex.
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影响因子：
10.5
作者：
LaBaer, J;Garrett, MD;Harlow, E
通讯作者：
Harlow, E
影响因子：
4.7
作者：
He G;Kuang J;Khokhar AR;Siddik ZH
通讯作者：
Siddik ZH
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
2.9
作者：
Adkins, JN;Lumb, KJ
通讯作者：
Lumb, KJ
影响因子：
8
作者：
Chopin, V;Toillon, RA;Le Bourhis, X
通讯作者：
Le Bourhis, X