Stracker Travis H, Morales Monica, Couto Suzana S, Hussein Hussein, Petrini John H J
Molecular Biology and Genetics, Sloan-Kettering Institute, New York, New York 10021, USA.
Nature. 2007 May 10;447(7141):218-21. doi: 10.1038/nature05740. Epub 2007 Apr 11.
The MRE11 complex (MRE11, RAD50 and NBS1) and the ataxia-telangiectasia mutated (ATM) kinase function in the same DNA damage response pathway to effect cell cycle checkpoint activation and apoptosis. The functional interaction between the MRE11 complex and ATM has been proposed to require a conserved C-terminal domain of NBS1 for recruitment of ATM to sites of DNA damage. Human Nijmegen breakage syndrome (NBS) cells and those derived from multiple mouse models of NBS express a hypomorphic NBS1 allele that exhibits impaired ATM activity despite having an intact C-terminal domain. This indicates that the NBS1 C terminus is not sufficient for ATM function. We derived Nbs1(DeltaC/DeltaC) mice in which the C-terminal ATM interaction domain is deleted. Nbs1(DeltaC/DeltaC) cells exhibit intra-S-phase checkpoint defects, but are otherwise indistinguishable from wild-type cells with respect to other checkpoint functions, ionizing radiation sensitivity and chromosome stability. However, multiple tissues of Nbs1(DeltaC/DeltaC) mice showed a severe apoptotic defect, comparable to that of ATM- or CHK2-deficient animals. Analysis of p53 transcriptional targets and ATM substrates showed that, in contrast to the phenotype of Chk2(-/-) mice, NBS1(DeltaC) does not impair the induction of proapoptotic genes. Instead, the defects observed in Nbs1(DeltaC/DeltaC) result from impaired phosphorylation of ATM targets including SMC1 and the proapoptotic factor, BID.
MRE11复合物(MRE11、RAD50和NBS1)与共济失调毛细血管扩张症突变(ATM)激酶在同一DNA损伤反应途径中发挥作用,以实现细胞周期检查点激活和细胞凋亡。有人提出,MRE11复合物与ATM之间的功能相互作用需要NBS1保守的C末端结构域,以便将ATM募集到DNA损伤位点。人类尼曼-匹克氏病(NBS)细胞以及源自多种NBS小鼠模型的细胞表达一种低表达的NBS1等位基因,尽管其C末端结构域完整,但ATM活性受损。这表明NBS1的C末端不足以实现ATM功能。我们培育出了缺失C末端ATM相互作用结构域的Nbs1(DeltaC/DeltaC)小鼠。Nbs1(DeltaC/DeltaC)细胞表现出S期内检查点缺陷,但在其他检查点功能、电离辐射敏感性和染色体稳定性方面与野生型细胞没有区别。然而,Nbs1(DeltaC/DeltaC)小鼠的多个组织表现出严重的凋亡缺陷,与ATM或CHK2缺陷动物的缺陷相当。对p53转录靶点和ATM底物的分析表明,与Chk2(-/-)小鼠的表型不同,NBS1(DeltaC)不会损害促凋亡基因的诱导。相反,在Nbs1(DeltaC/DeltaC)中观察到的缺陷是由于包括SMC1和促凋亡因子BID在内的ATM靶点磷酸化受损所致。