Wade R, Feldman D, Gunning P, Kedes L
Department of Medicine, Stanford Medical School, Palo Alto, CA.
Mol Cell Biochem. 1989 Jun 1;87(2):119-36. doi: 10.1007/BF00219255.
In order to initiate the study of the functional differences between myosin alkali light chain isoforms and to investigate the mechanisms of their differential expression, we have isolated cDNA clones for two human alkali light chain isoforms. Here we report DNA sequence and RNA blotting analyses that demonstrate that these cDNAs represent transcripts encoding human MLC3F and MLC1Sb. The sequence of the human MLC1Sb cDNA offers the first fully characterized example of a slow-fiber skeletal muscle alkali light chain isoform from any species. The sequence analysis of these two cDNAs allows an examination of evolutionarily conserved features of mammalian alkali light chain genes. Examination of the genomic organization of the human alkali light chain isoform genes revealed that, in contrast with some strains of mice, both are single copy genes. RNA blot analysis conclusively demonstrates that the human skeletal muscle MLC1Sb gene is also expressed in the heart ventricle but not the atria. In addition, we examined the expression of alkali light chain isoforms during the in vitro differentiation of a variety of human and rodent myogenic cells and found striking variation in the pattern of alkali light chain isoform gene expression in different myogenic cells.
为了启动对肌球蛋白碱轻链同工型功能差异的研究并探究其差异表达的机制,我们分离了两个人类碱轻链同工型的cDNA克隆。在此,我们报告DNA序列和RNA印迹分析,结果表明这些cDNA代表编码人类MLC3F和MLC1Sb的转录本。人类MLC1Sb cDNA的序列提供了来自任何物种的慢肌纤维骨骼肌碱轻链同工型的首个完全表征的实例。对这两个cDNA的序列分析使得能够检查哺乳动物碱轻链基因的进化保守特征。对人类碱轻链同工型基因的基因组组织检查表明,与某些品系的小鼠不同,二者均为单拷贝基因。RNA印迹分析确凿地证明,人类骨骼肌MLC1Sb基因也在心室而非心房中表达。此外,我们检查了多种人类和啮齿类成肌细胞体外分化过程中碱轻链同工型的表达,发现在不同成肌细胞中碱轻链同工型基因表达模式存在显著差异。