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J. Biol. Chem., Vol. 255, Issue 8, 3230-3233, 04, 1980

In vitro synthesis of heat-shock proteins by mRNAs from chicken embryo fibroblasts

PM Kelley, G Aliperti and MJ Schlesinger

The pattern of proteins synthesized by chicken embryo fibroblasts changes dramatically after these cells are incubated at 45 degrees C for a few hours. Three proteins (Mr = 22,000, 76,000, and 95,000) account for almost 50% of the cell's protein synthetic capacity immediately after the heat-shock (Kelley, P.M., and Schlesinger, M.J. (1978) Cell 15, 1277-1286). When mRNAs were isolated from heat-shocked cells and translated in a cell-free protein synthesizing system, a pattern of proteins virtually identical with that made by intact heat- shocked cells was detected. Mobilities in sodium dodecyl sulfate- polyacrylamide gel electrophoresis and radioimmune precipitation with specific antisera were used to establish the identity of in vitro- and in vivo-generated heat-shock proteins. The mRNAs coding for the major heat-shock proteins could be separated by rate zonal centrifugation in a sucrose gradient and mRNAs with sedimentation coefficients of 20 S, 18 S, and 13 S were translated in vitro to yield proteins of 95, 76, and 22 kilodaltons, respectively.
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				R. Currie and F. White Trauma-induced protein in rat tissues: a physiological role for a "heat shock" protein? Science, October 2, 1981; 214(4516): 72 - 73. [Abstract] [PDF]