Subramaniam M, Shanmugam G
J Cell Physiol. 1986 Jan;126(1):47-52. doi: 10.1002/jcp.1041260107.
Gene expression in quiescent mouse embryo fibroblasts was studied by labelling the cells with [14C] amino acids and analysing the proteins by electrophoresis in polyacrylamide gradient gels containing sodium dodecyl sulfate. Cycloheximide (CH) pretreatment of the cells was found to induce the synthesis of four proteins of molecular weights 72,000, 68,000, 42,000, and 29,000. These proteins were induced by CH both in serum-arrested and serum-stimulated cells. Addition of platelet-derived growth factor to serum-arrested quiescent cells also induced the synthesis of these proteins. Addition of CH and fetal calf serum (20%) to quiescent cells resulted in a dramatic increase in the synthesis of actin and another protein of molecular weight 29,000. The 29,000-dalton protein was present in higher quantities in the nuclei of induced cells. This protein appeared to be an early protein whose synthesis was transiently induced in quiescent cells within 3 hours of addition of 20% fetal calf serum (FCS). The synthesis of this protein was virtually turned off at 5-6 hours after the addition of serum. However, if CH or a combination of CH and FCS was present, a continuous synthesis of the 29 K protein was observed.
通过用[14C]氨基酸标记细胞,并在含十二烷基硫酸钠的聚丙烯酰胺梯度凝胶中进行电泳分析蛋白质,研究了静止小鼠胚胎成纤维细胞中的基因表达。发现用环己酰亚胺(CH)预处理细胞可诱导合成分子量分别为72,000、68,000、42,000和29,000的四种蛋白质。这些蛋白质在血清停滞和血清刺激的细胞中均由CH诱导产生。向血清停滞的静止细胞中添加血小板衍生生长因子也可诱导这些蛋白质的合成。向静止细胞中添加CH和胎牛血清(20%)导致肌动蛋白和另一种分子量为29,000的蛋白质的合成显著增加。29,000道尔顿的蛋白质在诱导细胞的细胞核中含量更高。这种蛋白质似乎是一种早期蛋白质,在添加20%胎牛血清(FCS)后3小时内,在静止细胞中短暂诱导合成。添加血清后5 - 6小时,这种蛋白质的合成几乎停止。然而,如果存在CH或CH与FCS的组合,则观察到29K蛋白质的持续合成。