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ATM regulates ionizing radiation-induced disruption of HDAC1 : PP1 : Rb complexes

机译:ATM调节电离辐射诱导的HDAC1:PP1:Rb复合物的破坏

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Ionizing radiation elicits signaling events that coordinate DNA repair and interruption of cell cycle progression. We previously demonstrated that ionizing radiation (IR) of cells activates nuclear protein phosphatase-1 (PP1) by promoting dephosphorylation of Thr320, an inhibitory site in the enzyme and that the ATM kinase is required for this response. We sought to identify potential targets of IR-activated PP1. Untreated and IR-treated Jurkat cells were labeled with P-32 orthophosphate, and nuclear extracts were subjected to microcystin affinity chromatography to recover phosphatase complexes that were analyzed by 2D-PAGE and mass spectrometry. Several proteins associated with protein phosphatases demonstrated a significant decrease in P-32 intensity following IR, and one of these was identified as HDAC1. Co-immunoprecipitation revealed complexes containing PP1 with HDAC1 and Rb in cell extracts. In response to IR, there was an ATM-dependent activation of PP1, dephosphorylation of HDAC1, dissociation of HDAC1-PP1-Rb complexes and increased HDAC1 activity. These results suggest that IR regulates HDAC1 phosphorylation and activity through ATM-dependent activation of PP1. (c) 2006 Elsevier Inc. All rights reserved.
机译:电离辐射引发协调DNA修复和细胞周期进程中断的信号事件。我们以前证明了细胞的电离辐射(IR)通过促进Thr320的去磷酸化(该酶的抑制位点)来激活核蛋白磷酸酶1(PP1),并且该反应需要ATM激酶。我们试图确定红外激活PP1的潜在目标。用P-32正磷酸盐标记未经处理和经IR处理的Jurkat细胞,然后对核提取物进行微囊藻毒素亲和层析,以回收磷酸酶复合物,并通过2D-PAGE和质谱分析。与蛋白质磷酸酶相关的几种蛋白质显示IR后P-32强度显着降低,其中一种被鉴定为HDAC1。免疫共沉淀显示细胞提取物中含有PP1与HDAC1和Rb的复合物。对IR的响应是PP1的ATM依赖性激活,HDAC1的去磷酸化,HDAC1-PP1-Rb复合物的解离和HDAC1活性的增加。这些结果表明IR通过依赖ATM的PP1激活来调节HDAC1的磷酸化和活性。 (c)2006 Elsevier Inc.保留所有权利。

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