Poly(ADP-ribose)-mediated interplay of XPA and PARP1 leads to reciprocal regulation of protein function.
Poly(ADP-ribose)-mediated interplay of XPA and PARP1 leads to reciprocal regulation of protein function.
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DOI：
10.1111/febs.12885
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发表时间：
2014-08
期刊：
影响因子：
--
通讯作者：
Bürkle A
中科院分区：
文献类型：
--
作者：
Fischer JM;Popp O;Gebhard D;Veith S;Fischbach A;Beneke S;Leitenstorfer A;Bergemann J;Scheffner M;Ferrando-May E;Mangerich A;Bürkle A
聚腺苷二磷酸核糖（Poly（ADP‐ribose），PAR）是一种复杂且可逆的翻译后修饰，通过共价修饰或非共价结合靶蛋白来控制蛋白质的功能和定位。以前，我们和其他人表征了关键核苷酸切除修复（NER）蛋白XPA与PAR的非共价，高亲和力结合。在本研究中，我们解决这种相互作用的功能相关性。首先，我们证实了细胞聚（ADP-核糖基）化（PAR化）的药理学抑制会损害NER的功效。其次，我们证明了XPA-PAR相互作用是由高度保守的PAR结合基序内的特定碱性氨基酸介导的，该基序与XPA的DNA损伤结合蛋白2（DDB 2）和转录因子II H（TFIIH）相互作用结构域重叠。第三，生物化学研究揭示了PARP 1和XPA功能的相互调节，一方面，XPA-PAR相互作用降低了XPA的DNA结合亲和力，而另一方面，XPA本身强烈刺激PARP 1酶活性。第四，U2 OS细胞中的微辐射实验表明，PARP抑制改变了XPA-绿色荧光蛋白对激光诱导的DNA损伤位点的募集特性。总之，我们的研究结果表明，XPA和PARP 1以相互和PAR依赖的方式相互调节，可能作为NER期间两个因子时空调节的微调机制。
Poly(ADP‐ribose) (PAR) is a complex and reversible post‐translational modification that controls protein function and localization through covalent modification of, or noncovalent binding to target proteins. Previously, we and others characterized the noncovalent, high‐affinity binding of the key nucleotide excision repair (NER) protein XPA to PAR. In the present study, we address the functional relevance of this interaction. First, we confirm that pharmacological inhibition of cellular poly(ADP‐ribosyl)ation (PARylation) impairs NER efficacy. Second, we demonstrate that the XPA–PAR interaction is mediated by specific basic amino acids within a highly conserved PAR‐binding motif, which overlaps the DNA damage‐binding protein 2 (DDB2) and transcription factor II H (TFIIH) interaction domains of XPA. Third, biochemical studies reveal a mutual regulation of PARP1 and XPA functions showing that, on the one hand, the XPA–PAR interaction lowers the DNA binding affinity of XPA, whereas, on the other hand, XPA itself strongly stimulates PARP1 enzymatic activity. Fourth, microirradiation experiments in U2OS cells demonstrate that PARP inhibition alters the recruitment properties of XPA‐green fluorescent protein to sites of laser‐induced DNA damage. In conclusion, our results reveal that XPA and PARP1 regulate each other in a reciprocal and PAR‐dependent manner, potentially acting as a fine‐tuning mechanism for the spatio‐temporal regulation of the two factors during NER.
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影响因子：
14.9
作者：
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影响因子：
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作者：
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