The KYxxL motif in Rad17 protein is essential for the interaction with the 9-1-1 complex
The KYxxL motif in Rad17 protein is essential for the interaction with the 9-1-1 complex
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DOI：
10.1016/j.bbrc.2016.07.014
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发表时间：
2016-09-02
影响因子：
3.1
通讯作者：
Yamaguchi, Naoto
中科院分区：
文献类型：
--
作者：
Fukumoto, Yasunori;Ikeuchi, Masayoshi;Yamaguchi, Naoto
依赖ATR的DNA损伤检查点是针对紫外线照射和DNA复制压力的主要DNA损伤检查点。Rad17 - RFC和Rad9 - Rad1 - Hus1（9 - 1 - 1）复合物相互作用以促进ATR信号传导，然而，这种相互作用的确切调控机制尚未确定。在此，我们在Rad17蛋白的AAA +结构域中鉴定出一个保守的序列基序KYxxL，并证明该基序对于与9 - 1 - 1复合物的相互作用至关重要。我们还表明，在Rad17 KYxxL突变体中，紫外线诱导的Rad17磷酸化增加。这些数据表明，与9 - 1 - 1复合物的相互作用对于Rad17蛋白成为紫外线诱导磷酸化的有效底物不是必需的。我们的数据还提出了9 - 1 - 1复合物在Rad17磷酸化中起负调控作用的可能性。我们还表明，Rad17的核苷酸结合活性是其核定位所必需的。（C）2016爱思唯尔公司。保留所有权利。
ATR-dependent DNA damage checkpoint is the major DNA damage checkpoint against UV irradiation and DNA replication stress. The Rad17-RFC and Rad9-Radl-Hus1 (9-1-1) complexes interact with each other to contribute to ATR signaling, however, the precise regulatory mechanism of the interaction has not been established. Here, we identified a conserved sequence motif, KYxxL, in the AAA+ domain of Rad17 protein, and demonstrated that this motif is essential for the interaction with the 9-1-1 complex. We also show that UV-induced Rad17 phosphorylation is increased in the Rad17 KYxxL mutants. These data indicate that the interaction with the 9-1-1 complex is not required for Rad17 protein to be an efficient substrate for the UV-induced phosphorylation. Our data also raise the possibility that the 9-1-1 complex plays a negative regulatory role in the Rad17 phosphorylation. We also show that the nucleotide-binding activity of Rad17 is required for its nuclear localization. (C) 2016 Elsevier Inc. All rights reserved.