Rebbe N F, Hickman W S, Ley T J, Stafford D W, Hickman S
Washington University Medical Service, Veterans Affairs Medical Center, St. Louis, Missouri 63106.
J Biol Chem. 1989 Sep 5;264(25):15006-11.
In order to define the mechanisms responsible for the regulated expression of a human 90-kDa heat shock protein (HSP90) gene, we have determined the complete genomic sequence of the gene encoding HSP90 beta, including 1102 base pairs upstream of the transcription initiation site. This gene consists of 12 exons and 11 introns. The exons range in size from 99 to 396 base pairs and the introns range from 91 to 1433 base pairs. Analysis of the 5' promoter region revealed TATA and CAAT consensus sequences. Additionally, Sp1 factor binding sites and presumptive heat shock element consensus sequences were found surrounding the transcription initiation site. Three out of the six identified potential heat shock elements reside within the first intron. The ability of the promoter region to respond to heat shock was examined by constructing a plasmid containing the 5' end of the HSP90 beta gene driving a neomycin phosphotransferase reporter gene. This plasmid was used to stably transform human K562 erythroleukemia cells. With heat shock, the abundance of HSP-neo transcripts increased, demonstrating that the promoter region of this gene contains the cis-acting DNA elements required for the heat shock response.
为了确定人类90 kDa热休克蛋白（HSP90）基因调控表达的机制，我们测定了编码HSP90β的基因的完整基因组序列，包括转录起始位点上游1102个碱基对。该基因由12个外显子和11个内含子组成。外显子大小从99到396个碱基对不等，内含子大小从91到1433个碱基对不等。对5'启动子区域的分析揭示了TATA和CAAT共有序列。此外，在转录起始位点周围发现了Sp1因子结合位点和假定的热休克元件共有序列。六个已鉴定的潜在热休克元件中有三个位于第一个内含子内。通过构建一个含有驱动新霉素磷酸转移酶报告基因的HSP90β基因5'端的质粒，检测了启动子区域对热休克的反应能力。该质粒用于稳定转染人K562红白血病细胞。热休克时，HSP-neo转录本的丰度增加，表明该基因的启动子区域含有热休克反应所需的顺式作用DNA元件。
