Features of a Smad3 MH1-DNA complex -: Roles of water and zinc in DNA binding

被引:59
作者
Chai, JJ
Wu, JW
Yan, N
Massagué, J
Pavletich, NP
Shi, YG [1 ]
机构
[1] Princeton Univ, Dept Mol Biol, Lewis Thomas Lab, Princeton, NJ 08544 USA
[2] Mem Sloan Kettering Canc Ctr, Howard Hughes Med Inst, Cell Biol Program, New York, NY 10021 USA
[3] Mem Sloan Kettering Canc Ctr, Howard Hughes Med Inst, Cellular Biochem & Biophys Program, New York, NY 10021 USA
关键词
D O I
10.1074/jbc.C300134200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Smad family of proteins mediates transforming growth factor-beta signaling from cell membrane to the nucleus. In the nucleus, Smads serve as transcription factors by directly binding to specific DNA sequences and regulating the expression of ligand-response genes. A previous structural analysis, at 2.8-Angstrom resolution, revealed a novel DNA-binding mode for the Smad MH1 domain but did not allow accurate assignment of the fines features of protein-DNA interactions. The crystal structure of a Smad3 MH1 domain bound to a palindromic DNA sequence, determined at 2.4-Angstrom resolution, reveals a surprisingly important role for water molecules. The asymmetric placement of the DNA-binding motif (a conserved 11-residue beta-hairpin) in the major groove of DNA is buttressed by seven well ordered water molecules. These water molecules make specific hydrogen bonds to the DNA bases, the DNA phosphate backbones, and several critical Smad3 residues. In addition, the MH1 domain is found to contain a bound zinc atom using four invariant residues among Smad proteins, three cysteines and one histidine. Removal of the zinc atom results in compromised DNA binding activity. These results define the Smad MH1 domain as a zinc-coordinating module that exhibits unique DNA binding properties.
引用
收藏
页码:20327 / 20331
页数:5
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