由 ATM/ATR 嵌合体揭示的 ATR N 端结构域的功能。
Function of the ATR N-terminal domain revealed by an ATM/ATR chimera.
作者信息
Chen Xinping, Zhao Runxiang, Glick Gloria G, Cortez David
机构信息
Department of Biochemistry, Vanderbilt University, Pierce Avenue, Nashville, TN 37232, USA.
出版信息
Exp Cell Res. 2007 May 1;313(8):1667-74. doi: 10.1016/j.yexcr.2007.02.015. Epub 2007 Feb 27.
The ATM and ATR kinases function at the apex of checkpoint signaling pathways. These kinases share significant sequence similarity, phosphorylate many of the same substrates, and have overlapping roles in initiating cell cycle checkpoints. However, they sense DNA damage through distinct mechanisms. ATR primarily senses single stranded DNA (ssDNA) through its interaction with ATRIP, and ATM senses double strand breaks through its interaction with Nbs1. We determined that the N-terminus of ATR contains a domain that binds ATRIP. Attaching this domain to ATM allowed the fusion protein (ATM*) to bind ATRIP and associate with RPA-coated ssDNA. ATM* also gained the ability to localize efficiently to stalled replication forks as well as double strand breaks. Despite having normal kinase activity when tested in vitro and being phosphorylated on S1981 in vivo, ATM* is defective in checkpoint signaling and does not complement cellular deficiencies in either ATM or ATR. These data indicate that the N-terminus of ATR is sufficient to bind ATRIP and to promote localization to sites of replication stress.
ATM和ATR激酶在细胞周期检查点信号通路的顶端发挥作用。这些激酶具有显著的序列相似性,磷酸化许多相同的底物,并且在启动细胞周期检查点方面具有重叠的作用。然而,它们通过不同的机制感知DNA损伤。ATR主要通过与ATRIP的相互作用来感知单链DNA(ssDNA),而ATM则通过与Nbs1的相互作用来感知双链断裂。我们确定ATR的N端包含一个与ATRIP结合的结构域。将该结构域连接到ATM上,使得融合蛋白(ATM*)能够结合ATRIP并与RPA包被的ssDNA结合。ATM还获得了有效定位于停滞的复制叉以及双链断裂处的能力。尽管在体外测试时具有正常的激酶活性,并且在体内S1981位点被磷酸化,但ATM在检查点信号传导方面存在缺陷,并且不能弥补ATM或ATR的细胞缺陷。这些数据表明,ATR的N端足以结合ATRIP并促进其定位于复制应激位点。