Davidson S, Høj P, Gabriele T, Anderson R L
Peter MacCallum Cancer Institute, Melbourne, Victoria, Australia.
Mol Cell Biol. 1995 Feb;15(2):1071-8. doi: 10.1128/MCB.15.2.1071.
We have identified a murine B-cell lymphoma cell line, CH1, that has a much-diminished capacity to express increased levels of heat shock proteins in response to heat stress in vitro. In particular, these cells cannot synthesize the inducible 72-kDa heat shock protein (HSP72) which is normally expressed at high levels in stressed cells. We show here that CH1 fails to transcribe HSP72 mRNA after heat shock, even though the heat shock transcription factor, HSF, is activated correctly. After heat shock, HSF from CH1 is found in the nucleus and is phosphorylated, trimerized, and capable of binding the heat shock element. We propose that additional signals which CH1 cells are unable to transduce are normally required to activate hsp72 transcription in vitro. Surprisingly, we have found that when the CH1 cells are heated in situ in a mouse, they show normal expression of HSP72 mRNA and protein. Therefore, CH1 cells have a functional hsp72 gene which can be transcribed and translated when the cells are in an appropriate environment. A diffusible factor present in ascites fluid is capable of restoring normal HSP72 induction in CH1 cells. We conclude that as-yet-undefined factors are required for regulation of the hsp72 gene or, alternatively, that heat shock in vivo causes activation of hsp70 through a novel pathway which the defect in CH1 has exposed and which is distinct from that operating in vitro. This unique system offers an opportunity to study a physiologically relevant pathway of heat shock induction and to biochemically define effectors involved in the mammalian stress response.
我们鉴定出一种小鼠B细胞淋巴瘤细胞系CH1,该细胞系在体外对热应激作出反应时,表达增加水平热休克蛋白的能力大幅降低。特别是,这些细胞无法合成诱导型72 kDa热休克蛋白(HSP72),而在应激细胞中该蛋白通常高水平表达。我们在此表明,CH1细胞在热休克后无法转录HSP72 mRNA,尽管热休克转录因子HSF被正确激活。热休克后,CH1细胞的HSF存在于细胞核中,且发生了磷酸化、三聚化,并且能够结合热休克元件。我们提出,在体外激活hsp72转录通常还需要CH1细胞无法转导的其他信号。令人惊讶的是,我们发现当CH1细胞在小鼠体内原位受热时,它们显示出HSP72 mRNA和蛋白的正常表达。因此,CH1细胞具有功能性hsp72基因,当细胞处于合适环境时该基因可以转录和翻译。腹水中存在的一种可扩散因子能够恢复CH1细胞中正常的HSP72诱导。我们得出结论,hsp72基因的调控需要尚未明确的因子,或者,体内热休克通过一种新途径导致hsp70激活,CH1细胞中的缺陷暴露了该途径,且该途径与体外起作用的途径不同。这个独特的系统为研究热休克诱导的生理相关途径以及从生物化学角度确定参与哺乳动物应激反应的效应物提供了机会。