Two N-terminal self-association domains are required for the dominant negative transcriptional activity of WT1 Denys-Drash mutant proteins

被引:36
作者
Holmes, G
Boterashvili, S
English, M
Wainwright, B
Licht, J
Little, M
机构
[1] UNIV QUEENSLAND,CTR CELLULAR & MOL BIOL,ST LUCIA,QLD 4072,AUSTRALIA
[2] UNIV QUEENSLAND,DEPT BIOCHEM,ST LUCIA,QLD 4072,AUSTRALIA
[3] MT SINAI SCH MED,BROOKDALE CTR MOL BIOL,NEW YORK,NY
关键词
D O I
10.1006/bbrc.1997.6545
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
Patients with Denys-Drash syndrome (DDS) have been shown to be constitutionally heterozygous for mutations of the WT1 gene. Almost all DDS mutations inactivate or remove the DNA-binding zinc finger region of WT1 and the resulting mutant proteins appear to act in a dominant negative manner. This may occur via WT1 self-association, which has been shown to involve the first 180 amino acids. By creating a series of N-terminal deletions, we have further investigated WT1 self-association using a yeast di-hybrid system and an in vitro protein binding assay. Our results suggest that there are two distinct domains within the N-terminal region facilitating self-association, residing from amino acids 1-45 and 157-253. Co-transfection of WT1 with progressively shorter N-terminal constructs demonstrates that both of these sites are required for a dominant negative activity as assessed by activation of a reporter construct. (C) 1997 Academic Press.
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页码:723 / 728
页数:6
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