Miyazaki J, Jozaki M, Nakatani N, Watanabe T, Saba R, Nakada K, Hirabayashi T, Yonemura I
Institute of Biological Sciences, University of Tsukuba, Japan.
J Muscle Res Cell Motil. 1999 Oct;20(7):655-60. doi: 10.1023/a:1005504018059.
To elucidate the mechanism that produces enormous molecular diversity in troponin T (TnT) of fast skeletal muscle, we determined the 5'-half genomic sequence of the chicken fast muscle TnT gene. The sequence of ca. 16 kb included seven exons (exons 1, 2, 3, 4, w, 5, and 6), which have been reported previously and presumed by sequencing TnT cDNAs. Additionally we found six 15 nt and one 18 nt sequences in the region between exons 5 and 6 (i.e. the exon x region). They were encompassed by consensus splice donor and acceptor sites and preceded by putative branch sites, and designated herein as exons xa to xg. Our result shows that the sequence derived from exons x1, x2, and x3, the exons presumed previously by cDNA sequencing, is actually encoded by the seven exons xa to xg, establishing the precise gene structure in the exon x region. Based on our data, together with that on the 3'-half genomic sequence of the quail fast muscle TnT gene, we conclude that the avian fast skeletal muscle TnT gene includes 27 exons, 16 of which are alternatively spliced.
为阐明在快速骨骼肌肌钙蛋白T(TnT)中产生巨大分子多样性的机制,我们测定了鸡快肌TnT基因的5′端半基因组序列。约16 kb的序列包含7个外显子(外显子1、2、3、4、w、5和6),这些外显子先前已被报道,并通过对TnT cDNA测序推测得到。此外,我们在外显子5和6之间的区域(即外显子x区域)发现了6个15 nt和1个18 nt的序列。它们被共有剪接供体和受体位点包围,且前面有推定的分支位点,在此处被命名为外显子xa至xg。我们的结果表明,先前通过cDNA测序推测的外显子x1、x2和x3的序列实际上由7个外显子xa至xg编码,从而确定了外显子x区域精确的基因结构。基于我们的数据,以及鹌鹑快肌TnT基因3′端半基因组序列的数据,我们得出结论,鸟类快速骨骼肌TnT基因包含27个外显子,其中16个可选择性剪接。