CHARACTERIZATION OF A MAMMALIAN SMOOTH-MUSCLE MYOSIN HEAVY-CHAIN GENE - COMPLETE NUCLEOTIDE AND PROTEIN CODING SEQUENCE AND ANALYSIS OF THE 5' END OF THE GENE
CHARACTERIZATION OF A MAMMALIAN SMOOTH-MUSCLE MYOSIN HEAVY-CHAIN GENE - COMPLETE NUCLEOTIDE AND PROTEIN CODING SEQUENCE AND ANALYSIS OF THE 5' END OF THE GENE
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DOI：
10.1073/pnas.88.23.10676
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发表时间：
1991-12-01
影响因子：
11.1
通讯作者：
PERIASAMY, M
中科院分区：
文献类型：
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作者：
BABIJ, P;KELLY, C;PERIASAMY, M
本研究的目的是表征编码兔平滑肌肌球蛋白重链（MHC）的完整cDNA序列，并确定相应基因5'端的外显子/内含子组织。从两个cDNA克隆PBRUC 1（几乎等于6.3个酶）和PBRU-PCR 33（420个碱基对）中产生了编码1972个氨基酸的蛋白质的全长cDNA序列，PBRUC 1（几乎等于6.3个酶）从兔子宫cDNA文库分离，PBRU-PCR 33（420个碱基对）通过引物延伸和PCR扩增产生。与鸡平滑肌MHC序列[Yanagisawa，M.，Hamada，Y.，Katsuragawa，Y.，Imamura，M.，Mikawa，T. & Masaki，T.等人（1987）J. Mol. 198，143-157]，兔MHC在S1球状头部区域中共享约90%的氨基酸同一性，但在功能上重要的S1头部结构域的25-kDa和50-kDa蛋白水解片段之间的连接处显示出显著的序列差异。基因组克隆表明，兔平滑肌MHC基因是大的，并在5'端有一个不寻常的外显子/内含子组织。前八个连续外显子位于基因组DNA的至少70个内切酶的区域内。一些内含子跨越DNA的几个内切酶，并且当与更远相关的肌节MHC基因相比时，5'端的其他内含子在Mg 2 +-ATP酶结构域中显示出高度的内含子保守性。引物延伸和S1核酸酶图谱分析表明，在兔平滑肌MHC基因的转录起始于一个单一的网站。
The purpose of this study was to characterize the complete cDNA sequence encoding the rabbit smooth muscle myosin heavy chain (MHC) and determine the exon/intron organization at the 5' end of the corresponding gene. The full-length cDNA sequence of 6644 base pairs encoding a protein of 1972 amino acids was generated from two cDNA clones: PBRUC1 (almost-equal-to 6.3 kilobases), isolated from a rabbit uterus cDNA library, and PBRU-PCR33 (420 base pairs), produced by primer extension and PCR amplification. Compared with the chicken smooth muscle MHC sequence [Yanagisawa, M., Hamada, Y., Katsuragawa, Y., Imamura, M., Mikawa, T. & Masaki, T. (1987) J. Mol. Biol. 198, 143-157] the rabbit MHC shares about 90% amino acid identity in the S1 globular head region but shows a striking sequence divergence at the junction between the 25-kDa and 50-kDa proteolytic fragments of the functionally important S1 head domain. Genomic cloning shows that the rabbit smooth muscle MHC gene is large and has an unusual exon/intron organization at the 5' end. The first eight contiguous exons are located within a region of at least 70 kilobases of genomic DNA. Some introns span several kilobases of DNA and others at the 5' end show a high degree of intron conservation in the Mg2+-ATPase domain when compared with more distantly related sarcomeric MHC genes. Primer extension and S1 nuclease mapping analysis demonstrate that transcription initiates from a single site in the rabbit smooth muscle MHC gene.