In vitro structure-function analysis of the beta-strand 326-333 of human p53

被引:33
作者
Chene, P [1 ]
Mittl, P [1 ]
Grutter, M [1 ]
机构
[1] NOVARTIS, CORE DRUG DISCOVERIES TECHNOL, CH-4002 BASEL, SWITZERLAND
关键词
p53; protein-protein interactions; beta-sheet; tetramerisation; protein interface;
D O I
10.1006/jmbi.1997.1360
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 beta-strand 326-333 is a key structural element in the formation of p53 tetramers. To investigate the contribution of its amino acid residues, an alanine scan was performed. The oligomerisation and DNA-binding properties of the mutant proteins were compared with those of wild-type proteins in vitro and analysed on the basis of the crystal structure of the p53 tetramerisation domain at 1.5 Angstrom resolution. Two categories of mutant proteins were identified. Phe328Ala, Leu330Ala and Ile332Ala mutant proteins are inactive for DNA binding and oligomerisation, while the Glu326Ala, Tyr327Ala, Thr329Ala, Gln331Ala and Arg333Ala mutant proteins have properties similar to those of wild-type proteins. These results suggest that single mutations within the p53 tetramerisation domain destabilise the structure of the whole protein, inhibiting its DNA-binding activity. Furthermore, the mutation of leucine 330 to alanine within the tetramerisation domain of the Arg175His protein abolishes the dominant negative effect of this mutant. This shows that the beta-strand 326-333 is a key structural element that mediates the dominant negative effect of p53 mutants. (C) 1997 Academic Press Limited.
引用
收藏
页码:873 / 881
页数:9
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