BRCA2 mediates centrosome cohesion via an interaction with cytoplasmic dynein
BRCA2 mediates centrosome cohesion via an interaction with cytoplasmic dynein
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DOI：
10.1080/15384101.2016.1195531
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发表时间：
2016-01-01
期刊：
影响因子：
4.3
通讯作者：
Nakanishi, Akira
中科院分区：
文献类型：
--
作者：
Malik, Sadiya;Saito, Hiroko;Nakanishi, Akira
BRCA 2与家族性乳腺癌和卵巢癌有关，并与DNA修复和中心体复制有关。在这里，我们分析了BRCA 2的中心体定位信号（CLS）与细胞质动力蛋白1相互作用，使BRCA 2定位于中心体的机制。体外拉下试验表明，BRCA 2直接结合细胞质动力蛋白1轻中间链2。显性负性HA-CLS-DsRed融合蛋白，通过siRNA耗尽动力蛋白，以及通过EHNA失活动力蛋白，抑制BRCA 2在中心体的定位，并导致S期中心体对分离。BRCA 2和C-Nap 1的双重缺失比C-Nap 1的沉默引起更大的中心体距离分散。这些结果表明，细胞质动力蛋白1结合BRCA 2通过后者的CLS和BRCA 2介导的凝聚力中心体之间在S期，潜在地作为一个细胞周期检查点。
BRCA2 is responsible for familial breast and ovarian cancer and has been linked to DNA repair and centrosome duplication. Here we analyzed the mechanism by which the centrosomal localization signal (CLS) of BRCA2 interacts with cytoplasmic dynein 1 to localize BRCA2 to the centrosome. In vitro pull-down assays demonstrated that BRCA2 directly binds to the cytoplasmic dynein 1 light intermediate chain 2. A dominant-negative HA-CLS-DsRed fusion protein, the depletion of dynein by siRNA, and the inactivation of dynein by EHNA, inhibited the localization of BRCA2 at centrosomes and caused the separation of centrosome pairs during the S-phase. The double depletion of BRCA2 and C-Nap1 caused a larger dispersion of centrosome distances than the silencing of C-Nap1. These results suggest that cytoplasmic dynein 1 binds to BRCA2 through the latter's CLS and BRCA2 mediates the cohesion between centrosomes during the S phase, potentially serving as a cell-cycle checkpoint.