Cloning and in situ hybridization of type 2A and 2B rat skeletal muscle myosin tail region: implications for filament assembly.
Cloning and in situ hybridization of type 2A and 2B rat skeletal muscle myosin tail region: implications for filament assembly.
复制标题
2A 型和 2B 型大鼠骨骼肌肌球蛋白尾区的克隆和原位杂交：对丝组装的影响。
DOI：
10.1006/bbrc.1993.2620
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发表时间：
1993
影响因子：
3.1
通讯作者：
Ryan,AF
中科院分区：
文献类型：
--
作者：
Lieber,RL;Bodine,SC;Burkholder,TJ;Pierotti,DJ;Ryan,AF
快速肌球蛋白表达的变化在骨骼肌适应中起关键作用。两种快速肌球蛋白亚型，2A型和2B型，通常由快速肌纤维表达，但它们的序列尚未确定，以允许mRNA表达研究。通过PCR扩增来自骨骼肌mRNA的cDNA，克隆到TA克隆载体中，并测序，获得一套完整的大鼠骨骼肌肌球蛋白。通过对骨骼肌的原位杂交和使用单克隆抗体鉴定肌球蛋白来证明特异性。克隆了两个新的肌球蛋白序列：2A型肌球蛋白（3′端642 bp）和2B型肌球蛋白（3′端624 bp）。这个区域编码肌球蛋白分子中与控制肌丝装配有关的部分。这两种快速肌球蛋白在开放阅读框架内的同源性为88%，在氨基酸水平上的同源性为95%。基于这种同源性，在2A型和2B型之间的肌纤维类型转化期间不太可能发生选择性肌球蛋白丝组装。
Changes in fast myosin expression play a critical role in skeletal muscle adaptation. Two fast myosin isoforms, type 2A and type 2B, are commonly expressed by fast muscle fibers but their sequences have not been determined to allow mRNA expression studies. A complete set of rat skeletal muscle myosins was amplified by PCR of cDNAs derived from skeletal muscle mRNA, cloned in a TA cloning vector, and sequenced. Specificity was demonstrated byin situhybridization against skeletal muscle and myosin protein identification using monoclonal antibodies. Two novel sequences were cloned: A type 2A myosin which consisted of a 642 bp segment from the 3′ end and a type 2B myosin which consisted of a 624 bp segment also from the 3′ end. This region encodes that portion of the myosin molecule implicated in the control of filament assembly. The two fast myosins showed 88% homology in the open reading frame and 95% homology at the amino acid level. Based on this homology, it is unlikely that selective myosin filament assembly occurs during muscle fiber type transformation between type 2A and 2B.