MUTATED YEAST HEAT-SHOCK TRANSCRIPTION FACTOR EXHIBITS ELEVATED BASAL TRANSCRIPTIONAL ACTIVATION AND CONFERS METAL RESISTANCE

被引:53
作者
SEWELL, AK
YOKOYA, F
YU, W
MIYAGAWA, T
MURAYAMA, T
WINGE, DR
机构
[1] UNIV UTAH, HLTH SCI CTR, DEPT MED, SALT LAKE CITY, UT 84132 USA
[2] UNIV UTAH, HLTH SCI CTR, DEPT BIOCHEM, SALT LAKE CITY, UT 84132 USA
[3] EHIME UNIV, FAC SCI, DEPT BIOL, MATSUYAMA, EHIME 790, JAPAN
关键词
D O I
10.1074/jbc.270.42.25079
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
Cadmium-resistant Saccharomyces cerevisiae strain 301N exhibits high basal as well as cadmium-induced expression of the CUP1 metallothionein gene. Since regulation of CUP1 is usually restricted to copper ions, our goal was to identify the factor responsible for the high metallothionein levels in strain 301N. The gene responsible for the observed phenotype is a spontaneously mutated heat shock transcription factor gene (HSF1). A double, semidominant HSF1 mutant with substitutions at codons 206 and 256 within the DNA-binding domain of the heat shock factor (HSF) confers two phenotypes. The first phenotype is elevated transcriptional activity of the HSF mutant (HSF301), which results in constitutive thermotolerance. A second HSF301 phenotype is enhanced binding affinity for the heat shock element (HSE) within the CUP1 5'-sequences, resulting in high basal transcription of metallothionein. The CUP1 HSE is a minimal heat shock element containing only two perfectly spaced inverted repeats of the basic nGAAn block. Cells containing HSF301 are resistant to cadmium salts. The single R206S mutation is responsible for the high affinity binding to the CUP1 HSE. In addition, the R206S HSF substitution exhibits constitutive transcriptional activation from a consensus HSE (HSE2). The F256Y substitution in HSF attenuates the effects of R206S on the consensus HSE2, but not on the CUP1 HSE.
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页码:25079 / 25086
页数:8
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