Ayme A, Southgate R, Tissières A
J Mol Biol. 1985 Apr 20;182(4):469-75. doi: 10.1016/0022-2836(85)90233-5.
The promoter regions of the Drosophila melanogaster small heat-shock protein genes have been analysed in order to localize those sequences responsible for their heat-shock transcriptional inducibility. Different lengths of the 5' DNA sequences of these four genes were each fused individually to the Herpes simplex virus thymidine kinase (HSV-tk) transcription unit. These hybrid genes were constructed in a simian virus 40 recombinant vector for transfection in permissive monkey COS cells and tested for their heat-shock inducibility. The hsp22/HSV-tk and hsp26/HSV-tk fusion genes were found to be heat-inducible at 43 degrees C, giving rise to correctly initiated transcripts, but transcriptionally quiescent at 37 degrees C (control temperature). The hsp23 and hsp27 fusion gene constructs are, however, not heat-shock-inducible; no transcripts being detectable from hsp27/HSV-tk constructs at either temperature and all hsp23/HSV-tk clones being faithfully but constitutively expressed at low levels at both temperatures. By testing a series of 5' deletion mutants in hsp22/HSV-tk, a homologous sequence located adjacent to the TATA box in both the hsp22 and hsp26 genes was identified as being responsible for their heat-shock activation. This control element corresponds to the Pelham "consensus sequence", previously described for the Drosophila hsp70 genes. The possible modes of transcriptional induction of all four genes are discussed.
为了定位那些负责果蝇小热休克蛋白基因热休克转录诱导性的序列,对其启动子区域进行了分析。这四个基因的不同长度的5' DNA序列分别单独与单纯疱疹病毒胸苷激酶(HSV-tk)转录单元融合。这些杂合基因构建在猴病毒40重组载体中,用于转染允许的猴COS细胞,并测试其热休克诱导性。发现hsp22/HSV-tk和hsp26/HSV-tk融合基因在43℃时可被热诱导,产生正确起始的转录本,但在37℃(对照温度)时转录静止。然而,hsp23和hsp27融合基因构建体不能被热休克诱导;在任何一个温度下,从hsp27/HSV-tk构建体中都检测不到转录本,并且所有hsp23/HSV-tk克隆在两个温度下都以低水平稳定但组成性地表达。通过测试hsp22/HSV-tk中的一系列5'缺失突变体,在hsp22和hsp26基因中与TATA框相邻的一个同源序列被确定为负责它们的热休克激活。这个控制元件对应于佩勒姆“共有序列”,先前已在果蝇hsp70基因中描述过。讨论了所有四个基因转录诱导的可能模式。