Malhotra S B, Bag J
Memorial University of Newfoundland, Faculty of Medicine, St. John's, Canada.
Mol Biol Rep. 1987;12(2):93-102. doi: 10.1007/BF00368876.
Cardiac myocyte cell culture from fourteen day old embryonic chicken heart was prepared. This cultured cell system was used to examine the regulation of troponin C (TnC) synthesis in cardiac muscle. To examine the regulation of TnC polypeptide synthesis, cardiac myocyte cells were pulse labelled with 35S-methionine at different days after plating. The synthesis of TnC was measured by determining the amount of radioactivity incorporated into the TnC polypeptide following separation by two dimensional gel electrophoresis. These measurements showed that TnC synthesis was maximum in 36 to 48 h old cultures and reached its lowest level in 4 day old cultures. This was in contrast to the synthesis of actin and tropomyosin. Synthesis of these polypeptides were lowest in 36 to 48 h old cultures and was maximum in 7 day old cultures. To examine whether the synthesis of TnC polypeptide paralleled the levels of TnC mRNA the sequences homologous to quail slow TnC cDNA clone were measured by hybridisation. The results showed that the decrease in the synthesis of troponin C polypeptide cannot be fully explained by the decrease in the steady state level of troponin C mRNA. The possibility of a role of translational control of troponin C mRNA in this process is discussed.
制备了来自14日龄胚胎鸡心脏的心肌细胞培养物。该培养细胞系统用于研究心肌中肌钙蛋白C(TnC)合成的调控。为了研究TnC多肽合成的调控,在接种后的不同天数用35S-甲硫氨酸对心肌细胞进行脉冲标记。通过二维凝胶电泳分离后,测定掺入TnC多肽中的放射性量来测量TnC的合成。这些测量结果表明,TnC合成在培养36至48小时时达到最大值,在培养4天时达到最低水平。这与肌动蛋白和原肌球蛋白的合成情况相反。这些多肽的合成在培养36至48小时时最低,在培养7天时最高。为了研究TnC多肽的合成是否与TnC mRNA水平平行,通过杂交测量与鹌鹑慢TnC cDNA克隆同源的序列。结果表明,肌钙蛋白C多肽合成的减少不能完全由肌钙蛋白C mRNA稳态水平的降低来解释。本文讨论了在此过程中TnC mRNA翻译控制作用的可能性。