Sreedharan R, Riordan M, Thullin G, Van Why S, Siegel N J, Kashgarian M
Medical College of Wisconsin, Wauwatosa, WI, USA.
Biochim Biophys Acta. 2011 Jan;1813(1):129-35. doi: 10.1016/j.bbamcr.2010.08.012. Epub 2010 Oct 8.
Endogenous heat shock proteins (HSPs) 70 and 25/27 are induced in renal cells by injury from energy depletion. Transfected over-expression of HSPs 70 or 27 (human analogue of HSP25), provide protection against renal cell injury from ATP deprivation. This study examines whether over-expressed HSP27 depends on induction of endogenous HSPs, in particular HSP70, to afford protection against cell injury. LLC-PK1 cells transfected with HSP27 (27OE cells) were injured by ATP depletion for 2h and recovered for 4h in the presence of HSF decoy, HSP70 specific siRNA (siRNA-70) and their respective controls. Injury in the presence of HSF decoy, a synthetic oligonucleotide identical to the heat shock element, the nuclear binding site of HSF, decreased HSP70 induction by 80% without affecting the over-expression of transfected HSP27. The HSP70 stress response was completely ablated in the presence of siRNA-70. Protection against injury, provided by over-expression of HSP27, was reduced by treatment with HSF decoy and abolished by treatment with siRNA-70. Immunoprecipitation studies demonstrated association of HSP27 with actin that was not affected by either treatment with HSF decoy or siRNA. Therefore, HSP27 is dependent on HSP70 to provide its maximal cytoprotective effect, but not for its interaction with actin. This study suggests that, while it has specific action on the cytoskeleton, HSP 25/27 must have coordinated activity with other HSP classes, especially HSP70, to provide the full extent of resistance to injury from energy depletion.
内源性热休克蛋白(HSPs)70和25/27可在肾细胞中由能量耗竭损伤诱导产生。转染过表达HSPs 70或27(HSP25的人类类似物)可提供针对ATP剥夺所致肾细胞损伤的保护作用。本研究探讨过表达的HSP27是否依赖内源性HSPs(尤其是HSP70)的诱导来提供针对细胞损伤的保护作用。用HSP27转染的LLC-PK1细胞(27OE细胞)经ATP剥夺损伤2小时,并在热休克因子(HSF)诱饵、HSP70特异性小干扰RNA(siRNA-70)及其各自对照存在的情况下恢复4小时。HSF诱饵是一种与热休克元件(HSF的核结合位点)相同的合成寡核苷酸,在其存在下,HSP70的诱导减少80%,而不影响转染的HSP27的过表达。在siRNA-70存在下,HSP70应激反应被完全消除。HSP27过表达所提供的针对损伤的保护作用,经HSF诱饵处理后降低,经siRNA-70处理后消除。免疫沉淀研究表明,HSP27与肌动蛋白的结合不受HSF诱饵或siRNA处理的影响。因此,HSP27依赖HSP70来提供其最大的细胞保护作用,但不依赖于其与肌动蛋白的相互作用。本研究表明,虽然HSP 25/27对细胞骨架有特定作用,但它必须与其他HSP类别(尤其是HSP70)协同发挥作用,以提供对能量耗竭损伤的全面抗性。