DNA-PK target identification reveals novel links between DNA repair signaling and cytoskeletal regulation.
DNA-PK target identification reveals novel links between DNA repair signaling and cytoskeletal regulation.
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DOI：
10.1371/journal.pone.0080313
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Quanz M
中科院分区：
文献类型：
--
作者：
Kotula E;Faigle W;Berthault N;Dingli F;Loew D;Sun JS;Dutreix M;Quanz M
DNA 依赖性蛋白激酶 (DNA-PK) 可能在 DNA 修复以外的各种细胞途径中充当关键信号激酶。使用二维凝胶电泳方法和稳定的 DNA 双链断裂模拟分子 (Dbait32Hc) 激活细胞核和细胞质中的 DNA-PK，我们鉴定了 26 种在 DNA-PK 激活后高度磷酸化的蛋白质。大多数这些蛋白质参与蛋白质稳定性和降解、细胞信号传导和细胞骨架。我们研究了 DNA-PK 与细胞骨架之间的关系，发现中间丝 (IF) 波形蛋白是体外和细胞内 DNA-PK 的靶标。在用 Dbait32Hc 转染的细胞中，波形蛋白通过 DNA-PK 在 Ser459 位点被磷酸化。我们制备了特异性抗体，结果表明 Ser459-P-vimentin 主要位于细胞突起处。在迁移细胞中，Dbait32Hc 诱导的波形蛋白磷酸化与较低的细胞粘附和迁移能力相关。因此，这种方法导致了 DNA-PK 下游细胞质靶标的鉴定，并揭示了 DNA 损伤信号传导与细胞骨架之间的联系。
The DNA-dependent protein kinase (DNA-PK) may function as a key signaling kinase in various cellular pathways other than DNA repair. Using a two-dimensional gel electrophoresis approach and stable DNA double-strand break-mimicking molecules (Dbait32Hc) to activate DNA-PK in the nucleus and cytoplasm, we identified 26 proteins that were highly phosphorylated following DNA-PK activation. Most of these proteins are involved in protein stability and degradation, cell signaling and the cytoskeleton. We investigated the relationship between DNA-PK and the cytoskeleton and found that the intermediate filament (IF) vimentin was a target of DNA-PK in vitro and in cells. Vimentin was phosphorylated at Ser459, by DNA-PK, in cells transfected with Dbait32Hc. We produced specific antibodies and showed that Ser459-P-vimentin was mostly located at cell protrusions. In migratory cells, the vimentin phosphorylation induced by Dbait32Hc was associated with a lower cellular adhesion and migration capacity. Thus, this approach led to the identification of downstream cytoplasmic targets of DNA-PK and revealed a connection between DNA damage signaling and the cytoskeleton.
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影响因子：
13.8
作者：
Flynn, Rachel Litman;Zou, Lee
通讯作者：
Zou, Lee
影响因子：
4.4
作者：
Garg, Ankita;Barnes, Peter F.;Vankayalapati, Ramakrishna
通讯作者：
Vankayalapati, Ramakrishna
影响因子：
4.3
作者：
Britton, Sebastien;Froment, Carine;Calsou, Patrick
通讯作者：
Calsou, Patrick
DOI：
10.1006/bbrc.1998.8741
发表时间：
1998-06-09
影响因子：
3.1
作者：
Basu, S;Rosenzweig, KR;Price, BD
通讯作者：
Price, BD
影响因子：
3.5
作者：
Athanasiou, A;Perunovic, B;Love, S
通讯作者：
Love, S