Regulation of hsp70 induction in thermotolerant HeLa cells.
Upon exposure to heat shock, non-thermotolerant (NT) HeLa cells transiently synthesize a large amount of 70-kDa heat-shock protein (hsp70), whereas thermotolerant (TT) cells synthesize a small amount of hsp70. When the hsp70 mRNA of HeLa cells was analyzed, it became apparent that hsp70 mRNA in TT cells did not increase following heat shock, whereas hsp70 mRNA in NT cells did increase dramatically. A further analysis of the activation of the heat-shock transcription factor (HSF) showed that significant activation of HSF was observed immediately after heat shock in both NT and TT cells. However, activated HSF was rapidly repressed in the TT cells, but not in the NT cells. Thus, the decreased induction of hsp70 synthesis observed in the TT HeLa cells may be due to the immediate repression of activated cellular HSF, which probably results in the reduced induction of hsp70 mRNA. The hsp70 content in the TT cells was usually higher than in the NT cells. However, after heat-shock treatment, the hsp70 content of the NT cells increased to nearly the level of the TT cells concomitant with the repression of hsp70 synthesis. The association of activated HSF with hsp70 was observed in both NT and TT cells, and the amount of HSF-hsp70 complex within the cell increased in proportion to the increase in hsp70 in the cells. These findings strongly suggest that the activity of HSF is negatively regulated by the intracellular content of hsp70 in these cells. Furthermore, in vitro experiments on the activation of HSF suggest that HSFs of NT and TT cells may have different properties, or an unknown factor may exist which regulates HSF activation in these cells.
Hatayama
T
T
Department of Biochemistry, Kyoto Pharmaceutical University, Japan.
Asai
Y
Y
Wakatsuki
T
T
Kitamura
T
T
Imahara
H
H
D001483:Base Sequence; D002457:Cell Extracts; D005786:Gene Expression Regulation; D006367:HeLa Cells / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis; D014157:Transcription Factors / Q000378:metabolism
D002457:Cell Extracts; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D014157:Transcription Factors