Nuclear focal adhesion kinase induces APC/C activator protein CDH1-mediated cyclin-dependent kinase 4/6 degradation and inhibits melanoma proliferation.
Nuclear focal adhesion kinase induces APC/C activator protein CDH1-mediated cyclin-dependent kinase 4/6 degradation and inhibits melanoma proliferation.
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DOI：
10.1016/j.jbc.2022.102013
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发表时间：
2022-06
影响因子：
4.8
通讯作者：
Lim, Ssang-Taek Steve
中科院分区：
文献类型：
--
作者：
Murphy, James M.;Jeong, Kyuho;Ahn, Eun-Young Erin;Lim, Ssang-Taek Steve
细胞周期蛋白依赖性激酶 (CDK) 的失调会促进不受控制的细胞增殖和癌症进展。尽管粘着斑激酶（FAK）有助于调节细胞周期进程，但确切的分子机制仍不清楚。在这里，我们发现 FAK 可能通过调节 CDK4/6 蛋白表达在细胞周期进程中发挥关键作用。我们发现 FAK 抑制增加了 B16F10 黑色素瘤细胞的核定位并诱导 G1 期停滞。从机制上讲，我们证明了核 FAK 与 CDK4/6 相关，并通过招募 CDC 同源物 1 (CDH1)（后期促进复合物/环体 E3 连接酶复合物的激活剂和底物识别亚基）促进其泛素化和蛋白酶体降解。我们发现 FAK N 端 FERM 结构域充当支架，使 CDK4/6 和 CDH1 紧密相连。然而，非核定位突变体 FAK FERM 的过度表达无法充当 CDK4/6 和 CDH1 的支架。此外，CDH1 的 shRNA 敲除增加了 B16F10 细胞中 CDK4/6 蛋白的表达，并阻止了 FAK 抑制剂诱导的 CDK4/6 的减少。在体内，我们发现与载体对照相比，药理学 FAK 抑制可减少 B16F10 肿瘤大小，这与 FAK 核定位增加和 CDK4/6 表达减少相关。在与患者匹配的健康皮肤和黑色素瘤活检中，我们发现健康皮肤中的 FAK 大部分处于非活性状态并且核位于健康皮肤中，而黑色素瘤病变则显示活性细胞质 FAK 增加和 CDK4 表达升高。总而言之，我们的数据表明，FAK 抑制通过诱导 G1 期停滞来阻止肿瘤增殖，部分是通过核 FAK 降低 CDK4/6 蛋白稳定性。
Dysregulation of cyclin-dependent kinases (CDKs) can promote unchecked cell proliferation and cancer progression. Although focal adhesion kinase (FAK) contributes to regulating cell cycle progression, the exact molecular mechanism remains unclear. Here, we found that FAK plays a key role in cell cycle progression potentially through regulation of CDK4/6 protein expression. We show that FAK inhibition increased its nuclear localization and induced G1 arrest in B16F10 melanoma cells. Mechanistically, we demonstrate nuclear FAK associated with CDK4/6 and promoted their ubiquitination and proteasomal degradation through recruitment of CDC homolog 1 (CDH1), an activator and substrate recognition subunit of the anaphase-promoting complex/cyclosome E3 ligase complex. We found the FAK N-terminal FERM domain acts as a scaffold to bring CDK4/6 and CDH1 within close proximity. However, overexpression of nonnuclear-localizing mutant FAK FERM failed to function as a scaffold for CDK4/6 and CDH1. Furthermore, shRNA knockdown of CDH1 increased CDK4/6 protein expression and blocked FAK inhibitor–induced reduction of CDK4/6 in B16F10 cells. In vivo, we show that pharmacological FAK inhibition reduced B16F10 tumor size, correlating with increased FAK nuclear localization and decreased CDK4/6 expression compared with vehicle controls. In patient-matched healthy skin and melanoma biopsies, we found FAK was mostly inactive and nuclear localized in healthy skin, whereas melanoma lesions showed increased active cytoplasmic FAK and elevated CDK4 expression. Taken together, our data demonstrate that FAK inhibition blocks tumor proliferation by inducing G1 arrest, in part through decreased CDK4/6 protein stability by nuclear FAK.
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影响因子：
10.5
作者：
Diehl, JA;Zindy, F;Sherr, CJ
通讯作者：
Sherr, CJ
影响因子：
16
作者：
Lim, Ssang-Taek;Chen, Xiao Lei;Llic, Dusko
通讯作者：
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影响因子：
158.5
作者：
Larkin, J.;Chiarion-Sileni, V.;Wolchok, J. D.
通讯作者：
Wolchok, J. D.
影响因子：
--
作者：
REISMAN, D;THOMPSON, EA
通讯作者：
THOMPSON, EA
DOI：
10.1083/jcb.201109067
发表时间：
2012-06-25
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lim ST;Miller NL;Chen XL;Tancioni I;Walsh CT;Lawson C;Uryu S;Weis SM;Cheresh DA;Schlaepfer DD
通讯作者：
Schlaepfer DD