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Mol Cell Biol. 1990 September; 10(9): 4886-4898

Induction of a chicken small heat shock (stress) protein: evidence of multilevel posttranscriptional regulation.

B V Edington and L E Hightower

Department of Molecular and Cell Biology, University of Connecticut, Storrs 06269.

ABSTRACT

A novel form of regulation of expression of a vertebrate heat shock gene is described. A cDNA clone encoding human Hsp27 was shown to specifically recognize chicken Hsp23 RNA by Northern (RNA) blot analysis and hybrid-select translation. This probe was then used to measure chicken hsp23 gene activity in control and heat-stressed cells. The hsp23 gene(s) was transcriptionally active in non-heat-stressed cells, and its rate of transcription did not increase significantly upon heat shock. Cytoplasmic Hsp23 mRNA, which was metabolically very stable in nonstressed cells, underwent a fourfold increase in amount after a 1-h heat shock, resulting in a twofold increase in Hsp23 mRNA in polysomes. Hsp23 mRNA was relatively abundant and translationally active even in non-heat-shocked cells. Taken together, these data implicated posttranscriptional nuclear events as an important control point for induction of Hsp23 RNA transcripts. The protein half-life of Hsp23 increased from approximately 2 h in control cultures to 13 h in heat-shocked cells, revealing a second major control point. Hsp23 which was synthesized prior to heat shock also increased in stability and contributed to the overall accumulation of Hsp23 in heat-shocked cells. Cycloheximide had no effect on this change in Hsp23 half-life, while dactinomycin blocked the stabilization of Hsp23, suggesting a need for newly synthesized RNA. These data indicated that stabilization of Hsp23 protein and posttranscriptional nuclear events resulting in increased production of Hsp23 mRNA were primarily responsible for a 13-fold increase in the accumulation of newly synthesized Hsp23 after 1 h of heat shock. The regulation of the hsp23 gene is discussed in comparison with several other posttranscriptionally regulated genes, including the proto-oncogene c-fos, the developmentally regulated chicken delta-crystallin gene, and regulation of cellular gene expression by the proto-oncogene c-myc.


Mol Cell Biol. 1990 September; 10(9): 4886-4898




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