CREATING NEW DNA-BINDING SPECIFICITIES IN THE YEAST TRANSCRIPTIONAL ACTIVATOR GCN4 BY COMBINING SELECTED AMINO-ACID SUBSTITUTIONS

被引：20

作者：

SUCKOW, M ^{[1
]}

MADAN, A ^{[1
]}

KISTERSWOIKE, B ^{[1
]}

VONWILCKENBERGMANN, B ^{[1
]}

MULLERHILL, B ^{[1
]}

机构：

[1] UNIV COLOGNE,INST GENET,D-50931 COLOGNE,GERMANY

来源：

NUCLEIC ACIDS RESEARCH | 1994年 / 22卷 / 12期

关键词：

D O I：

10.1093/nar/22.12.2198

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 specificity of the GCN4/DNA complex is mediated by a complicated network of interactions between the basic regions of both GCN4 monomers and their target halfsites. According to X-ray analyses (1, 2) one particular thymine of the target sequence is recognized by serine -11 and alanine -15 (we define the leucine in the first d-position of the heptad repeats as +1). We replaced serine -11 or alanine -15 with all other amino acids and analysed the DNA binding properties of the resulting stable GCN4 derivatives by electrophoretic mobility shift assays. Among these, mutants with tryptophan in position -11, or glutamic acid and glutamine in position -15, differ significantly from GCN4 in their DNA binding specificities. We then constructed selected double mutants, which differ from GCN4 in positions -11, -15 or -14 (3) of the basic region. The double mutants with tryptophan in position -11 and asparagine or serine in position -14 show drastically altered DNA binding specificities, presumably due to additive effects.

引用

页码：2198 / 2208

页数：11

共 32 条

[1] COGNATE DNA-BINDING SPECIFICITY RETAINED AFTER LEUCINE ZIPPER EXCHANGE BETWEEN GCN4 AND C/EBP [J].