Morimoto R, Fodor E
J Cell Biol. 1984 Oct;99(4 Pt 1):1316-23. doi: 10.1083/jcb.99.4.1316.
We have found that chicken reticulocytes respond to elevated temperatures by the induction of only one heat shock protein, HSP70, whereas lymphocytes induce the synthesis of all four heat shock proteins (89,000 mol wt, HSP89; 70,000 mol wt, HSP70; 23,000 mol wt, HSP23; and 22,000 mol wt, HSP22). The synthesis of HSP70 in lymphocytes was rapidly induced by small increases in temperature (2 degrees-3 degrees C) and blocked by preincubation with actinomycin D. Proteins normally translated at control temperatures in reticulocytes or lymphocytes were not efficiently translated after incubation at elevated temperatures. The preferential translation of mRNAs that encode the heat shock proteins paralleled a block in the translation of other cellular proteins. This effect was most prominently observed in reticulocytes where heat shock almost completely repressed alpha- and beta-globin synthesis. HSP70 is one of the major nonglobin proteins in chicken reticulocytes, present in the non-heat-shocked cell at approximately 3 X 10(6) molecules per cell. We compared HSP70 from normal and heat-shocked reticulocytes by two-dimensional gel electrophoresis and by digestion with Staphylococcus aureus V8 protease and found no detectable differences to suggest that the P70 in the normal cell is different from the heat shock-induced protein, HSP70. P70 separated by isoelectric focusing gel electrophoresis into two major protein spots, an acidic P70A (apparent pl = 5.95) and a basic P70B (apparent pl = 6.2). We observed a tissue-specific expression of P70A and P70B in lymphocytes and reticulocytes. In lymphocytes, P70A is the major 70,000-mol-wt protein synthesized at normal temperatures whereas only P70B is synthesized at normal temperatures in reticulocytes. Following incubation at elevated temperatures, the synthesis of both HSP70A and HSP70B was rapidly induced in lymphocytes, but synthesis of only HSP70B was induced in reticulocytes.
我们发现,鸡网织红细胞在温度升高时仅诱导一种热休克蛋白HSP70的产生,而淋巴细胞则诱导所有四种热休克蛋白(分子量89,000,HSP89;分子量70,000,HSP70;分子量23,000,HSP23;分子量22,000,HSP22)的合成。淋巴细胞中HSP70的合成在温度小幅升高(2摄氏度 - 3摄氏度)时迅速被诱导,并被放线菌素D预孵育所阻断。在网织红细胞或淋巴细胞中,正常温度下正常翻译的蛋白质在高温孵育后不能有效翻译。编码热休克蛋白的mRNA的优先翻译与其他细胞蛋白质翻译的阻断平行。这种效应在网织红细胞中最为明显,在那里热休克几乎完全抑制了α-和β-珠蛋白的合成。HSP70是鸡网织红细胞中主要的非珠蛋白之一,在未受热休克的细胞中,每个细胞约有3×10⁶个分子。我们通过二维凝胶电泳以及用金黄色葡萄球菌V8蛋白酶消化,比较了正常和热休克网织红细胞中的HSP70,未发现可检测到的差异,这表明正常细胞中的P70与热休克诱导的蛋白HSP70没有区别。P70通过等电聚焦凝胶电泳分离成两个主要蛋白斑点,一个酸性的P70A(表观pI = 5.95)和一个碱性的P70B(表观pI = 6.2)。我们观察到P70A和P70B在淋巴细胞和网织红细胞中有组织特异性表达。在淋巴细胞中,P70A是正常温度下合成的主要分子量70,000的蛋白,而在网织红细胞中正常温度下仅合成P70B。在高温孵育后,淋巴细胞中HSP70A和HSP70B的合成均迅速被诱导,但网织红细胞中仅诱导了HSP70B的合成。