Katsuragawa Y, Yanagisawa M, Inoue A, Masaki T
Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki, Japan.
Eur J Biochem. 1989 Oct 1;184(3):611-6. doi: 10.1111/j.1432-1033.1989.tb15057.x.
Two distinct cDNA clones for nonmuscle myosin heavy chain (MHC) were isolated from a chicken fibroblast cDNA library by cross-hydridization under a moderate stringency with chicken gizzard smooth muscle MHC cDNA. These two fibroblast MHC and the gizzard MHC are each encoded in different genes in the chicken genome. Northern blot analysis showed that both of the nonmuscle MHC mRNAs were expressed not only in fibroblasts but also in a variety of tissues including brain, lung, kidney, spleen, and skeletal, cardiac and smooth muscles. However, the relative contents of the two nonmuscle MHC mRNAs varied greatly among tissues. The encoded amino acid sequences of the nonmuscle MHCs were highly similar to each other (81% identity) and to the smooth muscle MHC (81-84%), but much less similar to vertebrate skeletal muscle MHCs (38-41%) or to protista nonmuscle MHCs (35-36%). A phylogenic tree of MHC isoforms was constructed by calculating the similarity scores between these MHC sequences. An examination of the tree showed that the vertebrate sarcomeric (skeletal and cardiac) MHC isoforms are encoded in a very closely related multigene family, and that the vertebrate non-sarcomeric (smooth muscle and nonmuscle) MHC isoforms define a distinct, less conserved MHC gene family.
通过与鸡砂囊平滑肌肌球蛋白重链(MHC)cDNA在中等严谨度下进行交叉杂交,从鸡成纤维细胞cDNA文库中分离出两个不同的非肌肉MHC cDNA克隆。这两种成纤维细胞MHC和砂囊MHC在鸡基因组中分别由不同的基因编码。Northern印迹分析表明,两种非肌肉MHC mRNA不仅在成纤维细胞中表达,而且在包括脑、肺、肾、脾以及骨骼肌、心肌和平滑肌在内的多种组织中也有表达。然而,这两种非肌肉MHC mRNA的相对含量在不同组织中差异很大。非肌肉MHC编码的氨基酸序列彼此高度相似(同一性为81%),与平滑肌MHC也高度相似(81 - 84%),但与脊椎动物骨骼肌MHC的相似性要低得多(38 - 41%),与原生动物非肌肉MHC的相似性更低(35 - 36%)。通过计算这些MHC序列之间的相似性得分构建了MHC同工型的系统发育树。对该树的检查表明,脊椎动物肌节(骨骼肌和心肌)MHC同工型由一个密切相关的多基因家族编码,而脊椎动物非肌节(平滑肌和非肌肉)MHC同工型则定义了一个不同的、保守性较低的MHC基因家族。