McDermott P, Whitaker-Dowling P, Klein I
University of Pittsburgh School of Medicine, Pennsylvania 15260.
Exp Cell Res. 1987 Nov;173(1):183-92. doi: 10.1016/0014-4827(87)90344-2.
Contraction regulates the myosin content and the rate of myosin synthesis in cultured neonatal rat heart cells. To further explore the mechanism for this regulation we examined various parameters of RNA content and RNA synthesis in contacting versus noncontracting myocytes. While contraction stimulated myosin heavy chain (MHC) synthesis by 72% compared to that of KCl-arrested cells, simultaneous analyses of polysome profiles were no different under the two culture conditions. Incorporation of [3H]uridine monophosphate into cellular RNA revealed no change in the rate of total RNA or ribosomal subunits synthesis. In vitro translation of cellular RNA yielded similar incorporation of [35S]methionine into trichloroacetic acid precipitable protein. Specific transcription of the MHC gene was examined by dot-blot analysis and was unaltered by contraction. Northern blot analysis of the MHC sequences detected by a cDNA probe revealed an mRNA sequence corresponding to a molecular weight of approximately 30 S. These data suggest that RNA synthesis and RNA content are unaltered by contraction in cultured heart cells and therefore the changes in myosin synthesis may be mediated at a post-transcriptional control level.
收缩调节培养的新生大鼠心脏细胞中的肌球蛋白含量和肌球蛋白合成速率。为了进一步探究这种调节机制,我们检测了收缩型与非收缩型心肌细胞中RNA含量和RNA合成的各种参数。与用氯化钾阻滞的细胞相比,收缩刺激肌球蛋白重链(MHC)合成增加了72%,但在两种培养条件下多核糖体图谱的同步分析并无差异。[3H]单磷酸尿苷掺入细胞RNA显示,总RNA或核糖体亚基的合成速率没有变化。细胞RNA的体外翻译产生了类似的[35S]甲硫氨酸掺入三氯乙酸可沉淀蛋白的情况。通过斑点印迹分析检测MHC基因的特异性转录,发现其不受收缩影响。用cDNA探针检测的MHC序列的Northern印迹分析显示,有一个对应分子量约为30 S的mRNA序列。这些数据表明,培养的心脏细胞中的RNA合成和RNA含量不受收缩影响,因此肌球蛋白合成的变化可能是在转录后控制水平介导的。