Xiao H, Lis J T
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Mol Cell Biol. 1989 Apr;9(4):1746-53. doi: 10.1128/mcb.9.4.1746-1753.1989.
In contrast to the hsp70 gene, whose expression is normally at a very low level and increases by more than 2 orders of magnitude during heat shock, the hsp83 gene in Drosophila melanogaster is expressed at high levels during normal development and increases only severalfold in response to heat shock. Developmental expression of the hsp83 gene consists of a high level of tissue-general, basal expression and a very high level of expression in ovaries. We identified regions upstream of the hsp83 gene that were required for its developmental and heat shock-induced expression by assaying beta-galactosidase activity and mRNA levels in transgenic animals containing a series of 5' deletion and insertion mutations of an hsp83-lacZ fusion gene. Deletion of sequences upstream of the overlapping array of a previously defined heat shock consensus (HSC) sequence eliminated both forms of developmental expression of the hsp83 gene. As a result, the hsp83 gene with this deletion mutation was regulated like that of the hsp70 gene. Moreover, an in vivo polymer competition assay revealed that the overlapping HSC sequences of the hsp83 gene and the nonoverlapping HSC sequences of the hsp70 gene had similar affinities for the factor required for heat induction of the two heat shock genes. We discuss the functional similarity of hsp70 and hsp83 heat shock regulation in terms of a revised view of the heat shock-regulatory sequence.
与hsp70基因不同,其表达通常处于非常低的水平,在热休克期间增加超过2个数量级,而黑腹果蝇中的hsp83基因在正常发育期间高水平表达,仅在热休克时增加几倍。hsp83基因的发育表达包括高水平的组织普遍基础表达和在卵巢中的非常高水平的表达。我们通过检测含有hsp83 - lacZ融合基因的一系列5'缺失和插入突变的转基因动物中的β - 半乳糖苷酶活性和mRNA水平,鉴定了hsp83基因上游对其发育和热休克诱导表达所需的区域。删除先前定义的热休克共有序列(HSC)重叠阵列上游的序列消除了hsp83基因的两种发育表达形式。结果,具有这种缺失突变的hsp83基因的调控方式与hsp70基因相同。此外,体内聚合物竞争试验表明,hsp83基因的重叠HSC序列和hsp70基因的非重叠HSC序列对这两个热休克基因热诱导所需的因子具有相似的亲和力。我们根据对热休克调控序列的修正观点讨论了hsp70和hsp83热休克调控的功能相似性。