TEMPERATURE-DEPENDENT REGULATION OF A HETEROLOGOUS TRANSCRIPTIONAL ACTIVATION DOMAIN FUSED TO YEAST HEAT-SHOCK TRANSCRIPTION FACTOR
TEMPERATURE-DEPENDENT REGULATION OF A HETEROLOGOUS TRANSCRIPTIONAL ACTIVATION DOMAIN FUSED TO YEAST HEAT-SHOCK TRANSCRIPTION FACTOR
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DOI：
10.1128/mcb.12.3.1021
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发表时间：
1992-03-01
影响因子：
5.3
通讯作者：
FACKENTHAL, DL
中科院分区：
文献类型：
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作者：
BONNER, JJ;HEYWARD, S;FACKENTHAL, DL
酿酒酵母的热休克转录因子（HSF）经过翻译后修饰。 在低生长温度下，它对热休克基因转录的激活作用很弱；转移到高温后，它很容易激活转录。 为了阐明这种调节机制，我们构建了一系列 HSF-VP16 融合体，将 HSF DNA 结合结构域与单纯疱疹病毒 VP16 基因的强转录激活结构域连接起来。 用 VP16 替换内源性 C 端转录激活结构域会生成 HSF 衍生物，其表现出让人想起 HSF 本身的行为：正常生长温度下的转录激活活性较低，热休克后的转录激活活性较高。 因此，HSF 可以抑制异源 VP16 转录激活结构域的活性。 为了确定在低温下抑制活性所需的条件，我们从 HSF-VP16 融合物中删除了部分 HSF，以绘制调控域图谱。 我们还分离了将 HSF-VP16 融合物转化为组成型转录激活因子的点突变。 我们得出的结论是，HSF 的中心进化保守结构域（包括 DNA 结合和多聚化结构域）包含温度依赖性调节的主要决定因素。
The heat shock transcription factor (HSF) of the yeast Saccharomyces cerevisiae is posttranslationally modified. At low growth temperatures, it activates transription of heat shock genes only poorly; after shift to high temperatures, it activates transcription readily. In an effort to elucidate the mechanism of this regulation, we constructed a series of HSF-VP16 fusions that join the HSF DNA-binding domain to the strong transcriptional activation domain from the VP16 gene of herpes simplex virus. Replacement of the endogenous C-terminal transcriptional activation domain with that of VP16 generates an HSF derivative that exhibits behavior reminiscent of HSF itself: low transcriptional activation activity at normal growth temperature and high activity after heat shock. HSF can thus restrain the activity of the heterologous VP16 transcriptional activation domain. To determine what is required for repression of activity at low temperature, we deleted portions of HSF from this HSF-VP16 fusion to map the regulatory domain. We also isolated point mutations that convert the HSF-VP16 fusion into a constitutive transcriptional activator. We conclude that the central, evolutionarily conserved domain of HSF, encompassing the DNA-binding and multimerization domains, contains a major determinant of temperature-dependent regulation.