Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins
Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins
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DOI：
10.1074/jbc.m006520200
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发表时间：
2000-12-29
影响因子：
4.8
通讯作者：
Althaus, FR
中科院分区：
文献类型：
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作者：
Pleschke, JM;Kleczkowska, HE;Althaus, FR
聚（ADP-核糖）可能在所有多细胞生物中由聚（ADP-核糖）聚合酶（PARP）家族形成。PARP-1是该家族中最好理解的并且直到最近才知道的唯一成员，是一种DNA损伤信号蛋白，其催化其与多个、大尺寸的ADP-核糖聚合物的自修饰，所述ADP-核糖聚合物可含有多达200个残基和几个分支点。通过这些聚合物，PARP-1可以与其他蛋白质非共价相互作用并改变它们的功能。在这里，我们报告的发现聚（ADP-核糖）结合序列基序在几个重要的DNA损伤检查点蛋白。20个氨基酸的基序包含两个保守区域：（i）富含碱性氨基酸的簇和（ii）散布有碱性残基的疏水性氨基酸模式。使用丙氨酸扫描、聚合物印迹分析和光亲和标记的组合，我们鉴定了以下蛋白质中的聚（ADP-核糖）结合位点：p53、p21（CIP 1/WAF 1）着色性干皮病A组补充蛋白、MSH 6、DNA连接酶III、XRCC 1、DNA聚合酶epsilon、DNA-PKCS、Ku 70、NF-κ B、诱导型一氧化氮合酶、半胱氨酸蛋白酶激活的DNA酶和端粒酶。发现聚（ADP-核糖）结合基序与负责（i）蛋白质-蛋白质相互作用，（ii）DNA结合，（iii）核定位，（iv）核输出和（v）蛋白质降解的五个重要功能结构域重叠。因此，PARP可以通过聚（ADP-核糖）靶向特定的信号网络蛋白并调节其结构域功能。
Poly(ADP-ribose) is formed in possibly all multicellular organisms by a familiy of poly(ADP-ribose) polymerases (PARPs). PARP-1, the best understood and until recently the only known member of this family, is a DNA damage signal protein catalyzing its automodification with multiple, variably sized ADP-ribose polymers that may contain up to 200 residues and several branching points. Through these polymers, PARP-1 can interact noncovalently with other proteins and alter their functions. Here we report the discovery of a poly(ADP-ribose)-binding sequence motif in several important DNA damage checkpoint proteins. The 20-amino acid motif contains two conserved regions: (i) a cluster rich in basic amino acids and (ii) a pattern of hydrophobic amino acids interspersed with basic residues. Using a combination of alanine scanning, polymer blot analysis, and photoaffinity labeling, we have identified poly(ADP-ribose)-binding sites in the following proteins: p53, p21(CIP1/WAF1) xeroderma pigmentosum group A complementing protein, MSH6, DNA ligase III, XRCC1, DNA polymerase epsilon, DNA-PKCS, Ku70, NF-kappaB, inducible nitric-oxide synthase , caspase-activated DNase, and telomerase. The poly(ADP-ribose)-binding motif was found to overlap with five important functional domains responsible for (i) protein-protein interactions, (ii) DNA binding, (iii) nuclear localization, (iv) nuclear export, and (v) protein degradation. Thus, PARPs may target specific signal network proteins via poly(ADP-ribose) and regulate their domain functions.