Heat shock induces two distinct S6 protein kinase activities in quiescent mammalian fibroblasts.
The regulation of S6 kinase activity was used to monitor perturbations of intracellular signaling activity during heat shock of quiescent murine and human fibroblasts. Previous reports on exponentially growing insect and plant cells had indicated that 40S ribosomal protein S6 is dephosphorylated during heat shock; thus inhibition of S6 kinase activity by heat shock was anticipated in NIH 3T3 fibroblasts and human cells (HeLa, diploid embryonic fibroblasts MRC-5, and skin-derived fibroblasts). Unexpectedly, two distinct S6 protein kinases were activated in quiescent fibroblasts after heat exposure. One of the enzymes was partially purified by sequential column chromatography and was determined to be equivalent to the enzyme activated by serum and other growth factors, referred to here as pp70-S6 protein kinase. The other protein S6 kinase, pp90rsk, was identified by a specific immunoprecipitation assay. Monitoring both enzymatic activities during heat shock revealed a temporal pattern of activation that was reversed when compared to non-stressed, mitogen-stimulated cells. Finally, heat shock stimulated protein S6 phosphorylation in cultured, quiescent mammalian cells. These data demonstrate that specific protein kinases can be activated during heat shock, and that some early mitogenic signals may also participate in the response of cells to physiologic stress.
Jurivich
D A
DA
Department of Medicine, Northwestern University Medical School, Chicago, Illinois 60611.
Chung
J
J
Blenis
J
J
D000818:Animals; D001769:Blood; D002460:Cell Line; D003470:Culture Media; D004789:Enzyme Activation; D004790:Enzyme Induction; D006367:HeLa Cells; D006358:Hot Temperature; D006801:Humans; D007399:Interphase; D010766:Phosphorylation; D011494:Protein Kinases / Q000096:biosynthesis* / Q000378:metabolism; D038601:Ribosomal Protein S6; D019893:Ribosomal Protein S6 Kinases; D012269:Ribosomal Proteins / Q000378:metabolism; D015398:Signal Transduction
D003470:Culture Media; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D011494:Protein Kinases; D019893:Ribosomal Protein S6 Kinases