Sequential assignments of the 1H NMR resonances of Zn(II)2 and 113Cd(II)2 derivatives of the DNA-binding domain of the GAL4 transcription factor reveal a novel structural motif for specific DNA recognition.
Sequential assignments of the 1H NMR resonances of Zn(II)2 and 113Cd(II)2 derivatives of the DNA-binding domain of the GAL4 transcription factor reveal a novel structural motif for specific DNA recognition.
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GAL4 转录因子 DNA 结合域的 Zn(II)2 和 113Cd(II)2 衍生物的 1H NMR 共振序列分配揭示了特异性 DNA 识别的新结构基序。
DOI：
10.1021/bi00231a016
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发表时间：
1991
期刊：
影响因子：
2.9
通讯作者：
Coleman,JE
中科院分区：
文献类型：
--
作者：
Pan,T;Coleman,JE
Department of Molecular Biophysics and Biochemistry，Yale University，New Haven，Connecticut 06510 Received August 8，1990; Revised Mandarin pt Received December 10，1990摘要：GAL 4转录因子的DNA结合结构域，由62个N-末端氨基酸残基组成，表示为GAL 4（62*），含有新的Zn（II）2Cys 6或Cd（II）2Cys 6双核簇[Pan，T.，&科尔曼，JE（1990）美国国家科学院院刊。Acad. Sci. USA 87，2077]。特异性DNA识别需要位于该双核簇内以及C末端的残基。用DQF-COSY、中继COSY、双中继COSY和NOESY对Zn（II）2-和U_3Cd（II）_2GAL_4（62 ~*）进行了核磁共振序列归属。两个四面体金属结合位点的配体已被鉴定为一个金属离子的Cys 11、Cys 14、Cys 21和Cys 31，以及另一个金属离子的Cys 28、Cys 38、Cys 21和Cys 31，其中Cys 21和Cys 31作为两个金属离子之间共享的配体。在GAL 4（62*）结构内可以发现无螺旋，其由一系列转角组成以容纳金属簇，随后是不规则环和从残基42至60的转角，“特异性区域”，其序列对特异性DNA识别有重要贡献。在形成双核簇的残基和特异性区域内的几个残基之间观察到长距离NOE，表明后者被折叠到金属簇上。Zn（II）2Cys 6双核簇和特异性区域与DNA结合的要求揭示了GAL 4作为一类特异性DNA结合蛋白的成员，其使用新的结构基序识别特异性DNA序列。这类蛋白质的特异性DNA结合是通过使用包围Zn（II）2Cys 6双核簇的转角和环来实现的，而不是如先前描述的特异性DNA结合蛋白中所观察到的-螺旋或/3-链。
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06510 Received August 8, 1990; Revised Manuscript Received December 10, 1990 abstract: The DNA-binding domain of the GAL4 transcription factor, consisting of the 62 N-terminal amino acid residues and denoted GAL4 (62*), contains a novel Zn (II) 2Cys6 or Cd (II) 2Cys6 binuclear cluster [Pan, T., & Coleman, JE (1990) Proc. Natl. Acad. Sci. USA 87, 2077]. Specific DNA recognition requires residues located within as well as C terminal to this binuclear cluster. NMR sequential assignments have been carried out on Zn (II) 2-and U3Cd (II) 2GAL4 (62*) by using DQF-COSY, relayed COSY, double-relayed COSY, and NOESY. The ligands of the two tetrahedral metal-binding sites have been identified as Cys11, Cys14, Cys21, and Cys31 to one metal ion and Cys28, Cys38, Cys21, and Cys31 to the other metal ion with Cys21 and Cys31 as ligands shared between the two metal ions. No-helices can be found within the GAL4 (62*) structure, which consists of a series of turns to accommodate the metal cluster, followed by irregular loops and turns from residues 42 to 60, the “specificity region”, whose sequence contributes importantly to specific DNA recognition. Long-distance NOE’s are observed between residues forming the binuclear cluster and several residues within the specificity region, indicatingthat the latter is folded compactly onto the metal cluster. The requirement of the Zn (II) 2Cys6 binuclear cluster and the specificity region for binding to DNA reveals GAL4 as a member of a class of specific DNA-binding proteins using a new structural motif for the recognition of specific DNA sequences. Specific DNA binding by this class of proteins is achieved by use of turns and loops that enclose a Zn (II) 2Cys6 binuclear cluster, instead of-helices or/3-strands as observed in specific DNA-binding proteins described previously.