Adaptive exchange sustains cullin-RING ubiquitin ligase networks and proper licensing of DNA replication.
Adaptive exchange sustains cullin-RING ubiquitin ligase networks and proper licensing of DNA replication.
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DOI：
10.1073/pnas.2205608119
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发表时间：
2022-09-06
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
去核化酶Cop9信号体（CSN）调节cullin-RING泛素连接酶（CRL）的功能，所述CRL通过泛素介导的蛋白质如转录因子、细胞周期调节剂和信号转导蛋白的降解参与调节无数细胞功能。通过小分子抑制剂CSN 5i-3的CSN抑制对CRL功能具有不同的影响。当CSN被抑制时，一些CRL被深度灭活，而其他CRL几乎不受影响。因此，很难去卷积CSN抑制对细胞的影响。为了解开这个问题，我们使用了CRISPRi和CRISPRa筛选，并确定了参与DNA复制许可的两个贡献途径。这些知识可能用于指导选择最有可能对CSN5i-3做出反应的癌症患者。Cop9信号体（CSN）通过将泛素样蛋白NEDD 8从cullin亚基上解偶联来调节cullin-RING E3泛素连接酶（CRL）的功能。为了了解CSN功能对CRL网络和细胞增殖的生理影响，我们将定量质谱和全基因组CRISPR干扰（CRISPRi）和CRISPR激活（CRISPRa）筛选相结合，以确定在小分子CSN5i-3抑制CSN后调节细胞活力的因素。CRL组分和调节剂强烈调节CSN 5i-3的抗增殖作用，此外，我们发现涉及基因组完整性的两条途径，SCFFBX05-APC/C-GMNN和CUL4DTL-SETD8，其实质上有助于CSN抑制的毒性。我们的数据突出了CSN介导的NEDD8去缀合和CRL底物受体的适应性交换在维持CRL功能中的重要性，并提出了利用CSN抑制治疗癌症的方法。
The deneddylase Cop9 signalosome (CSN) regulates the function of cullin-RING ubiquitin ligases (CRLs), which are involved in regulating myriad cellular functions through the ubiquitin-mediated degradation of proteins such as transcription factors, cell cycle regulators, and signal transduction proteins. CSN inhibition by the small molecule inhibitor, CSN5i-3, has differential effects on CRL function. Some CRLs are profoundly inactivated when CSN is inhibited, whereas others are barely affected. Therefore, it is difficult to deconvolute the effects of CSN inhibition on cells. To untangle this, we used CRISPRi and CRISPRa screens and identified two contributing pathways involved in DNA replication licensing. The knowledge can potentially be used to guide selection of cancer patients that are most likely to respond to CSN5i-3. Cop9 signalosome (CSN) regulates the function of cullin–RING E3 ubiquitin ligases (CRLs) by deconjugating the ubiquitin-like protein NEDD8 from the cullin subunit. To understand the physiological impact of CSN function on the CRL network and cell proliferation, we combined quantitative mass spectrometry and genome-wide CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) screens to identify factors that modulate cell viability upon inhibition of CSN by the small molecule CSN5i-3. CRL components and regulators strongly modulated the antiproliferative effects of CSN5i-3, and in addition we found two pathways involved in genome integrity, SCFFBXO5–APC/C–GMNN and CUL4DTL–SETD8, that contribute substantially to the toxicity of CSN inhibition. Our data highlight the importance of CSN-mediated NEDD8 deconjugation and adaptive exchange of CRL substrate receptors in sustaining CRL function and suggest approaches for leveraging CSN inhibition for the treatment of cancer.
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影响因子：
--
作者：
Cope GA;Deshaies RJ
通讯作者：
Deshaies RJ
DOI：
10.1074/jbc.m112.352484
发表时间：
2012-08-24
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Emberley ED;Mosadeghi R;Deshaies RJ
通讯作者：
Deshaies RJ
影响因子：
4.8
作者：
Houston, Sabrina I.;McManus, Kirk J.;Rice, Judd C.
通讯作者：
Rice, Judd C.
影响因子：
16
作者：
Abbas T;Shibata E;Park J;Jha S;Karnani N;Dutta A
通讯作者：
Dutta A
影响因子：
8.8
作者：
Enchev RI;Scott DC;da Fonseca PC;Schreiber A;Monda JK;Schulman BA;Peter M;Morris EP
通讯作者：
Morris EP