USP1-trapping lesions as a source of DNA replication stress and genomic instability.
USP1-trapping lesions as a source of DNA replication stress and genomic instability.
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DOI：
10.1038/s41467-022-29369-3
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发表时间：
2022-04-01
影响因子：
16.6
通讯作者：
Huang TT
中科院分区：
文献类型：
--
作者：
Coleman KE;Yin Y;Lui SKL;Keegan S;Fenyo D;Smith DJ;Rothenberg E;Huang TT
去泛素化酶USP 1通过范可尼贫血蛋白和DNA复制持续性因子增殖细胞核抗原（PCNA）的去泛素化作用，是基因组完整性的关键调节因子。独特的是，在紫外线照射后，USP 1通过自切割自失活，这使得其自身降解，进而上调PCNA单泛素化。然而，在生理条件下，这种自裂解事件的功能作用仍然难以捉摸。在此，我们发现，细胞窝藏的自切割缺陷的USP 1突变体，同时仍然能够稳健地去泛素化PCNA，经历更多的复制叉停滞和提前叉终止事件。使用超分辨率显微镜和活细胞单分子跟踪，我们发现这些缺陷与这种USP 1突变体无法从活性DNA合成位点正确回收有关，从而导致复制相关病变。此外，我们还发现DNA依赖性金属蛋白酶Spartan可以促进USP 1分子从DNA中的去除，以抵消“USP 1捕获”引起的细胞毒性。我们提出了一个实用的USP 1抑制剂在癌症治疗中的基础上，他们的能力，诱导USP 1捕获病变和随之而来的复制压力和基因组不稳定性的癌细胞，类似于非共价的DNA-蛋白质交联如何通过施加空间位阻蛋白参与DNA交易引起细胞毒性。在这里，作者提供了关于USP 1去泛素化酶的自动切割如何调节DNA复制和基因组稳定性的机制见解。通过蛋白质-DNA捕获在癌症治疗中的USP 1活性的靶向的意义进行了讨论。
The deubiquitinase USP1 is a critical regulator of genome integrity through the deubiquitylation of Fanconi Anemia proteins and the DNA replication processivity factor, proliferating cell nuclear antigen (PCNA). Uniquely, following UV irradiation, USP1 self-inactivates through autocleavage, which enables its own degradation and in turn, upregulates PCNA monoubiquitylation. However, the functional role for this autocleavage event during physiological conditions remains elusive. Herein, we discover that cells harboring an autocleavage-defective USP1 mutant, while still able to robustly deubiquitylate PCNA, experience more replication fork-stalling and premature fork termination events. Using super-resolution microscopy and live-cell single-molecule tracking, we show that these defects are related to the inability of this USP1 mutant to be properly recycled from sites of active DNA synthesis, resulting in replication-associated lesions. Furthermore, we find that the removal of USP1 molecules from DNA is facilitated by the DNA-dependent metalloprotease Spartan to counteract the cytotoxicity caused by “USP1-trapping”. We propose a utility of USP1 inhibitors in cancer therapy based on their ability to induce USP1-trapping lesions and consequent replication stress and genomic instability in cancer cells, similar to how non-covalent DNA-protein crosslinks cause cytotoxicity by imposing steric hindrances upon proteins involved in DNA transactions. Here the authors provide mechanistic insights into how auto-cleavage of the USP1 deubiquitinase regulates DNA replication and genome stability. Implications for the targeting of USP1 activity via protein-DNA trapping in cancer therapy are discussed.
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影响因子：
64.8
作者：
Ray Chaudhuri A;Callen E;Ding X;Gogola E;Duarte AA;Lee JE;Wong N;Lafarga V;Calvo JA;Panzarino NJ;John S;Day A;Crespo AV;Shen B;Starnes LM;de Ruiter JR;Daniel JA;Konstantinopoulos PA;Cortez D;Cantor SB;Fernandez-Capetillo O;Ge K;Jonkers J;Rottenberg S;Sharan SK;Nussenzweig A
通讯作者：
Nussenzweig A
影响因子：
16
作者：
Békés M;van der Heden van Noort GJ;Ekkebus R;Ovaa H;Huang TT;Lima CD
通讯作者：
Lima CD
DOI：
10.1007/978-1-4939-1680-1_10
发表时间：
2015
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Dungrawala, Huzefa;Cortez, David
通讯作者：
Cortez, David
影响因子：
16
作者：
Cohn, Martin A.;Kowal, Przemyslaw;D'Andrea, Alan D.
通讯作者：
D'Andrea, Alan D.
影响因子：
10.5
作者：
Ge, Xin Quan;Jackson, Dean A;Blow, J Julian
通讯作者：
Blow, J Julian