Publication Date:
1988-12-16
Description:
A novel method has been developed for modulating the expression of an endogenous chromosomal gene in a higher eukaryote, by competitive inhibition at the level of gene transcription. The gene studied was the hsp70 gene, which encodes a 72-kilodalton (kD) heat shock protein that is synthesized after thermal stress. The 5' control region of the hsp70 gene was inserted on a plasmid containing the eukaryotic gene for dihydrofolate reductase. The hybrid plasmid was then introduced into a Chinese hamster ovary cell line and elevated in copy number approximately 20,000-fold by selection of cells with methotrexate. Heat-inducible expression from the intact hsp70 gene was reduced by at least 90% in the modified cells when compared with the induction in control cells, and the modified cells also displayed elevated thermosensitivity. The change in heat shock protein synthesis is presumably caused by competition among the increased number of binding sites for the heat-shock transcription factor, leading to altered expression from the native heat shock gene. These results support a role for heat shock protein in the recovery of mammalian cells from acute thermal stress.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Johnston, R N -- Kucey, B L -- New York, N.Y. -- Science. 1988 Dec 16;242(4885):1551-4.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biological Sciences, University of Calgary, Canada.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/3201244" target="_blank"〉PubMed〈/a〉
Keywords:
Animals
;
Cell Line
;
Cell Survival
;
DNA/genetics/isolation & purification
;
Gene Amplification
;
*Gene Expression Regulation
;
*Genes
;
Heat-Shock Proteins/biosynthesis/*genetics
;
Hot Temperature
;
Methotrexate/pharmacology
;
Tetrahydrofolate Dehydrogenase/genetics
;
*Transcription, Genetic
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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