Identification of chromatin-related protein interactions using protein microarrays.
Identification of chromatin-related protein interactions using protein microarrays.
复制标题
使用蛋白质微阵列鉴定染色质相关蛋白质相互作用。
DOI：
10.1002/pmic.200300593
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发表时间：
2003
期刊：
影响因子：
3.4
通讯作者：
Albala,JoannaS
中科院分区：
文献类型：
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作者：
Coleman,MatthewA;Miller,KristiA;Beernink,PeterT;Yoshikawa,DanielM;Albala,JoannaS
染色质中的动态结构变化由蛋白质相互作用介导，所述蛋白质相互作用调节多个细胞过程，包括复制、转录、重组和DNA修复。识别染色质的复合物是由几组不同的蛋白质定义的，这些蛋白质要么直接修饰组蛋白，要么与组蛋白-DNA复合物相互作用。我们将蛋白质、抗体和DNA应用于功能化的载玻片，并用我们感兴趣的蛋白质询问载玻片，以鉴定参与DNA双链断裂修复的蛋白质的新型蛋白质-蛋白质相互作用。在这里，我们证明了DNA修复蛋白RAD 51 B，而不是它的同源伙伴RAD 51 C，与组蛋白，而不是核小体相互作用。最近鉴定的SWI/SNF蛋白SMARCAL 1证明了核小体特异性相互作用。使用Far Western分析证实了独特的RAD 51 B-组蛋白相互作用。这是第一次证明RAD 51 B和组蛋白之间的相互作用，这可能对DNA双链断裂的成功修复很重要。
Dynamic structural changes in chromatin are mediated by protein interactions that modulate multiple cellular processes including replication, transcription, recombination and DNA repair. Complexes that recognize chromatin are defined by several distinct groups of proteins that either directly modify histones or interact with histone‐DNA complexes.A protein microarray format was used to analyze the interaction of various DNA repair proteins with chromatin components. We applied proteins, antibodies and DNA to functionalized glass slides and interrogated the slides with our proteins of interest to identify novel protein‐protein interactions for proteins involved in DNA double‐strand break repair. Here we demonstrate that the DNA repair protein RAD51B, and not its cognate partner RAD51C, interacts with histones and not nucleosomes. Nucleosome‐specific interactions were demonstrated with the recently identified SWI/SNF protein, SMARCAL1. Unique RAD51B‐histone interactions were corroborated using Far Western analysis. This is the first demonstration of an interaction between RAD51B and histone proteins that may be important for the successful repair of DNA double‐strand breaks.