Servant P, Mazodier P
Unité de Biochimie Microbienne, Institut Pasteur, Paris, France.
J Bacteriol. 1996 Dec;178(24):7031-6. doi: 10.1128/jb.178.24.7031-7036.1996.
In Streptomyces albus G, HSP18, a protein belonging to the small heat shock protein family, could be detected only at high temperature. The nucleotide sequence of the DNA region upstream from hsp18 contains an open reading frame (orfY) which is in the opposite orientation and 150 bp upstream. This open reading frame encodes a basic protein of 225 amino acids showing no significant similarity to any proteins found in data banks. Disruption of this gene in the S. albus chromosome generated mutants that synthesized hsp18 RNA at 30 degrees C, suggesting that orfY plays either a direct or indirect role in the transcriptional regulation of the hsp18 gene. In addition, thermally induced expression of the hsp18 gene is subject to posttranscriptional regulation. In the orfY mutant, although hsp18 RNA was synthesized at a high level at 30 degrees C, the HSP18 protein could not be detected except after heat shock. Synthesis of the HSP18 protein in the orfY mutant was also heat inducible when transcription was inhibited by rifampin. Furthermore, when wild-type cultures of S. albus were shifted from high temperature to 30 degrees C, synthesis of the gene product could no longer be detected, even though large amounts of hsp18 RNA were present.
在白色链霉菌G中,属于小热休克蛋白家族的HSP18仅在高温下才能被检测到。hsp18上游DNA区域的核苷酸序列包含一个开放阅读框(orfY),其方向相反,位于上游150 bp处。这个开放阅读框编码一个由225个氨基酸组成的碱性蛋白,与数据库中发现的任何蛋白质都没有明显的相似性。在白色链霉菌染色体中破坏这个基因会产生在30℃时合成hsp18 RNA的突变体,这表明orfY在hsp18基因的转录调控中起直接或间接作用。此外,hsp18基因的热诱导表达受到转录后调控。在orfY突变体中,尽管hsp18 RNA在30℃时大量合成,但除热休克后外,无法检测到HSP18蛋白。当利福平抑制转录时,orfY突变体中HSP18蛋白的合成也可被热诱导。此外,当白色链霉菌的野生型培养物从高温转移到30℃时,即使存在大量的hsp18 RNA,也无法再检测到基因产物的合成。