Croissant J D, Carpenter S, Bader D
Gladys P. Stahlman Cardiovascular Research Laboratory, Vanderbilt University Medical Center, Nashville, Tennessee 37212-6300, USA.
J Biol Chem. 2000 Jan 21;275(3):1944-51. doi: 10.1074/jbc.275.3.1944.
We report the identification and cloning of a unique chick myosin heavy chain (CMHC1) that is expressed exclusively in the heart during embryogenesis. Using primers specific to myosin heavy chains, we used reverse transcriptase-polymerase chain reaction to clone and isolate CMHC1 from embryonic day 10 chicken heart RNA. Sequence analysis indicated that CMHC1 was a novel member of the myosin heavy chain family. Expression of the CMHC1 transcripts was detected in Hamburger Hamilton stage 10 chick embryos in the fusing myocardium. Expression of CMHC1 was maintained at high levels throughout the tubular heart of later stage embryos. Reverse transcriptase-polymerase chain reaction and in situ hybridizations failed to detect CMHC1 transcripts in the developing somites, limb buds, or skeletal musculature at any stage of chick development. Genomic CMHC1 clones have been isolated that contain sequences approximately 5.2 kilobase upstream of the presumptive CMHC1 transcription start site. Portions of the upstream regulatory region induced a 21-fold increase in reporter gene expression in primary cardiomyocytes. Because of its unique cardiac-restricted expression, CMHC1 will provide an excellent model system to study the molecular mechanisms required for the early developmental regulation of heart-specific genes.
我们报告了一种独特的鸡肌球蛋白重链(CMHC1)的鉴定和克隆,该蛋白在胚胎发育过程中仅在心脏中表达。我们使用肌球蛋白重链特异性引物,通过逆转录聚合酶链反应从胚胎第10天的鸡心脏RNA中克隆并分离出CMHC1。序列分析表明,CMHC1是肌球蛋白重链家族的一个新成员。在融合心肌的汉伯格-汉密尔顿10期鸡胚中检测到CMHC1转录本的表达。在后期胚胎的管状心脏中,CMHC1的表达一直维持在高水平。逆转录聚合酶链反应和原位杂交未能在鸡发育的任何阶段的发育中的体节、肢芽或骨骼肌组织中检测到CMHC1转录本。已分离出基因组CMHC1克隆,其包含推定的CMHC1转录起始位点上游约5.2千碱基的序列。上游调控区的部分序列在原代心肌细胞中使报告基因表达增加了21倍。由于其独特的心脏限制性表达,CMHC1将为研究心脏特异性基因早期发育调控所需的分子机制提供一个优秀的模型系统。