Mulligan-Tuttle Anne, Heikkila John J
Department of Biology, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
Comp Biochem Physiol A Mol Integr Physiol. 2007 Oct;148(2):308-16. doi: 10.1016/j.cbpa.2007.04.022. Epub 2007 May 3.
In the present study, we examined the expression of the Rana catesbeiana small heat shock protein gene, hsp30, in an FT fibroblast cell line. Northern and western blot analyses revealed that hsp30 mRNA or HSP30 protein was not present constitutively but was strongly induced at a heat shock temperature of 35 degrees C. However, treatment of FT cells with sodium arsenite at concentrations that induced hsp gene expression in other amphibian systems caused cell death. Non-lethal concentrations of sodium arsenite (10 microM) induced only minimal accumulation of hsp30 mRNA or protein after 12 h. Immunocytochemical analyses employing laser scanning confocal microscopy detected the presence of heat-inducible HSP30, in a granular or punctate pattern. HSP30 was enriched in the nucleus with more diffuse localization in the cytoplasm. The nuclear localization of HSP30 was more prominent with continuous heat shock. These heat treatments did not alter FT cell shape or disrupt actin cytoskeletal organization. Also, HSP30 did not co-localize with the actin cytoskeleton.
在本研究中,我们检测了牛蛙小热休克蛋白基因hsp30在FT成纤维细胞系中的表达。Northern印迹和Western印迹分析显示,hsp30 mRNA或HSP30蛋白并非组成性存在,而是在35℃热休克温度下被强烈诱导表达。然而,在其他两栖动物系统中能诱导hsp基因表达的亚砷酸钠浓度处理FT细胞时,却导致细胞死亡。非致死浓度的亚砷酸钠(10μM)在12小时后仅诱导hsp30 mRNA或蛋白产生极少的积累。采用激光扫描共聚焦显微镜进行的免疫细胞化学分析检测到热诱导型HSP30以颗粒状或点状模式存在。HSP30在细胞核中富集,在细胞质中的定位更为弥散。持续热休克时,HSP30的核定位更为显著。这些热处理并未改变FT细胞的形状,也未破坏肌动蛋白细胞骨架的组织。此外,HSP30与肌动蛋白细胞骨架没有共定位。