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纯化的人因子在无细胞的HeLa转录系统中激活热休克启动子。

Purified human factor activates heat shock promoter in a HeLa cell-free transcription system.

作者信息

Goldenberg C J, Luo Y, Fenna M, Baler R, Weinmann R, Voellmy R

机构信息

Department of Microbiology and Immunology, University of Miami School of Medicine, Florida 33101.

出版信息

J Biol Chem. 1988 Dec 25;263(36):19734-9.

PMID:3198647
Abstract

Heat shock protein (hsp) genes are typically silent and are activated by various stresses including heat. As a first step toward understanding this activation event, a human factor, referred to here as human heat shock transcription factor (human HTF), has been purified approximately 14,000-fold from extracts of heat-treated HeLa cells by means of sequence-specific DNA affinity chromatography. The most highly purified fraction of human HTF binds specifically to the known regulatory sequence element (HSE) of hsp genes as shown by footprinting experiments. Purified human HTF has an apparent molecular mass of 83 kDa. Human HTF is specifically required for activation of an hsp gene promoter in a reconstituted in vitro transcription system from human cells. Activation is dependent on the presence of the HSEs in the transcription template.

摘要

热休克蛋白(hsp)基因通常处于沉默状态,可被包括热在内的各种应激激活。作为理解这一激活事件的第一步,一种人类因子,在此称为人类热休克转录因子(人类HTF),已通过序列特异性DNA亲和层析从热处理的HeLa细胞提取物中纯化了约14000倍。足迹实验表明,人类HTF的最高纯化级分与hsp基因已知的调控序列元件(HSE)特异性结合。纯化的人类HTF的表观分子量为83 kDa。在重组的人类细胞体外转录系统中,hsp基因启动子的激活特别需要人类HTF。激活依赖于转录模板中HSE的存在。

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