The protein-binding potential of C2H2 zinc finger domains
The protein-binding potential of C2H2 zinc finger domains
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DOI：
10.1007/s12013-008-9007-6
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发表时间：
2008-05-01
影响因子：
2.6
通讯作者：
Segal, David J.
中科院分区：
文献类型：
--
作者：
Brayer, Kathryn J.;Kulshreshtha, Sanjeev;Segal, David J.
在人类基因组中，有超过10000个c2h2型锌指（ZF）结构域分布在1000多个ZF蛋白中。这些结构域经常被观察到参与序列特异性DNA结合，而非特征结构域通常被认为是促进DNA相互作用。然而，一些zf也促进与蛋白质或RNA的结合。超过100种Cys2-His2 (C2H2) zf蛋白相互作用已被描述。我们最初尝试进行生物信息学分析，以确定序列特征，以预测DNA或蛋白质结合功能。这些努力由于几个问题而变得复杂，包括zf的全部功能的不确定性。因此，我们采用了一种无偏倚的方法来直接检查ZFs促进DNA或蛋白质相互作用的潜力。以人OLF-1/EBF相关锌指蛋白（OAZ）为模型。人类O/ e -1相关锌指蛋白（hOAZ）在6个簇中包含30个zf，其中一些已在DNA或蛋白质相互作用中被发现。DNA结合使用目标位点选择（CAST）试验评估，蛋白质结合使用酵母双杂交试验评估。我们观察到，已知结合DNA的簇可以促进特定的蛋白质相互作用，但已知结合蛋白质的簇不能促进特定的DNA相互作用。我们的主要结论是，DNA结合是ZFs更有限的功能，它们介导蛋白质相互作用的潜力可能更大。这些结果表明，C2H2 ZF结构域在蛋白质相互作用中的作用可能被低估了。讨论了这些发现对ZF函数预测的意义。
There are over 10,000 C2H2-type zinc finger (ZF) domains distributed among more than 1,000 ZF proteins in the human genome. These domains are frequently observed to be involved in sequence-specific DNA binding, and uncharacterized domains are typically assumed to facilitate DNA interactions. However, some ZFs also facilitate binding to proteins or RNA. Over 100 Cys2-His2 (C2H2) ZF-protein interactions have been described. We initially attempted a bioinformatics analysis to identify sequence features that would predict a DNA- or protein-binding function. These efforts were complicated by several issues, including uncertainties about the full functional capabilities of the ZFs. We therefore applied an unbiased approach to directly examine the potential for ZFs to facilitate DNA or protein interactions. The human OLF-1/EBF associated zinc finger (OAZ) protein was used as a model. The human O/E-1-associated zinc finger protein (hOAZ) contains 30 ZFs in 6 clusters, some of which have been previously indicated in DNA or protein interactions. DNA binding was assessed using a target site selection (CAST) assay, and protein binding was assessed using a yeast two-hybrid assay. We observed that clusters known to bind DNA could facilitate specific protein interactions, but clusters known to bind protein did not facilitate specific DNA interactions. Our primary conclusion is that DNA binding is a more restricted function of ZFs, and that their potential for mediating protein interactions is likely greater. These results suggest that the role of C2H2 ZF domains in protein interactions has probably been underestimated. The implication of these findings for the prediction of ZF function is discussed.