THE REDOX AND DNA-REPAIR ACTIVITIES OF REF-1 ARE ENCODED BY NONOVERLAPPING DOMAINS
THE REDOX AND DNA-REPAIR ACTIVITIES OF REF-1 ARE ENCODED BY NONOVERLAPPING DOMAINS
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DOI：
10.1073/pnas.91.1.23
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发表时间：
1994-01-04
影响因子：
11.1
通讯作者：
CURRAN, T
中科院分区：
文献类型：
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作者：
XANTHOUDAKIS, S;MIAO, GG;CURRAN, T
转录因子AP-1的DNA结合活性在体外由涉及还原/氧化（氧化还原）的翻译后机制调节。氧化还原调节是由一个保守的半胱氨酸残基介导的DNA结合域的Fos和Jun。以前，我们证明了DNA修复蛋白，参考-1，可以刺激DNA结合活性的Fos-Jun二聚体通过减少这个半胱氨酸残基。为了研究Ref-1的氧化还原和修复功能之间的关系，我们产生了一系列缺失突变体。在体外的截断蛋白的分析表明，氧化还原和修复活动的Ref-1的不同区域编码。Ref-1的N-末端结构域中不存在于来自其他生物的功能相关蛋白质中的序列是氧化还原活性所需的，而DNA修复活性需要保守的C-末端序列。半胱氨酸巯基的化学烷基化或氧化抑制Ref-1的氧化还原活性，而不影响其DNA修复活性。交联研究表明，直接半胱氨酸介导的相互作用之间发生Ref-1和Jun.
The DNA binding activity of transcription factor AP-1 is regulated in vitro by a posttranslational mechanism involving reduction/oxidation (redox). Redox regulation is mediated by a conserved cysteine residue in the DNA-binding domain of Fos and Jun. Previously, we demonstrated that a DNA repair protein, Ref-1, could stimulate the DNA binding activity of Fos-Jun dimers by reducing this cysteine residue. To examine the relationship between the redox and repair functions of Ref-1, we generated a series of deletion mutants. Analysis of the truncated proteins in vitro revealed that the redox and repair activities are encoded by distinct regions of Ref-1. Sequences in the N-terminal domain of Ref-1 that are not present in functionally related proteins from other organisms are required for the redox activity, whereas the DNA repair activity requires conserved C-terminal sequences. Chemical alkylation or oxidation of cysteine sulfhydryls inhibits the redox activity of Ref-1 without affecting its DNA repair activity. Crosslinking studies suggest that a direct cysteine-mediated interaction occurs between Ref-1 and Jun.