Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells.
Heat shock induces in cells the synthesis of specific proteins called heat shock proteins (HSPs) and a transient state of thermotolerance. The putative role of one of the HSPs, HSP27, as a protective molecule during thermal stress has been directly assessed by measuring the resistance to hyperthermia of Chinese hamster and mouse cells transfected with the human HSP27 gene contained in plasmid pHS2711. One- and two-dimensional gel electrophoresis of [3H]leucine- and [32P]orthophosphate-labeled proteins, coupled with immunological analysis using Ha27Ab and Hu27Ab, two rabbit antisera that specifically recognize the hamster and the human HSP27 protein respectively, were used to monitor expression and inducibility of the transfected and endogenous proteins. The human HSP27 gene cloned in pHS2711 is constitutively expressed in rodent cells, resulting in accumulation of the human HSP27 and all phosphorylated derivatives. No modification of the basal or heat-induced expression of endogenous HSPs is detected. The presence of additional HSP27 protein provides immediate protection against heat shock administered 48 h after transfection and confers a permanent thermoresistant phenotype to stable transfectant Chinese hamster and mouse cell lines. Mild heat treatment of the transfected cells results in an induction of the full complement of the endogenous heat shock proteins and a small increase in thermoresistance, but the level attained did not surpass that of heat-induced thermotolerant control cells. These results indicate that elevated levels of HSP27 is sufficient to give protection from thermal killing. It is concluded that HSP27 plays a major role in the increased thermal resistance acquired by cells after exposure to HSP inducers.
Landry
J
J
Centre de Recherche en Cancérologie, Université Laval, l'Hôtel-Dieu de Québec, Canada.
Chrétien
P
P
Lambert
H
H
Hickey
E
E
Weber
L A
LA
D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival; D002478:Cells, Cultured; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000235:genetics / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D010750:Phosphoproteins / Q000502:physiology; D014162:Transfection
D006360:Heat-Shock Proteins; D010750:Phosphoproteins