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单个鸡心肌肌球蛋白碱性轻链基因通过一个复杂外显子的可变剪接产生两种不同的mRNA。

Single chicken cardiac myosin alkali light-chain gene generates two different mRNAs by alternative splicing of a complex exon.

作者信息

Nakamura S, Nabeshima Y, Kobayashi H, Nabeshima Y, Nonomura Y, Fujii-Kuriyama Y

机构信息

Department of Biochemistry, Cancer Institute, Japanese Foundation for Cancer Research, Tokyo.

出版信息

J Mol Biol. 1988 Oct 20;203(4):895-904. doi: 10.1016/0022-2836(88)90115-5.

Abstract

We have isolated and characterized two kinds of cDNA for the chicken cardiac myosin alkali light chain. The sequences of the two cDNAs are identical, except for a notable divergence in part of the 3' untranslated sequence. By analysis of isolated genomic clones, it was shown that the genomic sequences corresponding to the different sequences in the 3' untranslated regions of the two mRNAs were arranged within a limited part of a single stretch of DNA; also the two distinct 3' untranslated regions of the two mRNAs shared part of the last exon, which was 0.6 x 10(3) base-pairs long. There are two canonical acceptor sites available for RNA splicing in the last exon, the first being located at the 5' end of the exon, and the second at 370 base-pairs downstream from this end. Together with analysis by S1 nuclease mapping, the foregoing results lead us to conclude that, by the differential use of these two acceptor sites, a single gene generates two distinct mRNAs of 1.45 x 10(3) base-pairs and 1.1 x 10(3) base-pairs with or without the 5' half of the last exon. The two mRNAs appear to utilize the same modified poly(A) signal, AGTAAA, rather than the authentic AATAAA sequence present about 30 base-pairs downstream from the poly(A) attachment sites. This is probably because another consensus G + T-rich sequence is present at an appropriate distance from the AGTAAA sequence, but not from the AATAAA sequence. The gene for the cardiac myosin alkali light chain has proved to be expressed in ventricular muscle and in atrial and anterior latissimus dorsi muscles, the last of these being characteristic of slow skeletal muscle. In these muscles, two kinds of mRNA for the cardiac myosin alkali light chain, identical with those in ventricular muscle, were expressed and their relative amount in each tissue was almost the same as that in ventricular muscle.

摘要

我们已分离并鉴定了鸡心肌肌球蛋白碱性轻链的两种cDNA。这两种cDNA的序列相同,只是3'非翻译序列的部分区域存在显著差异。通过对分离的基因组克隆进行分析,结果表明,对应于两种mRNA的3'非翻译区不同序列的基因组序列排列在一段单一DNA的有限区域内;两种mRNA不同的3'非翻译区还共享最后一个外显子的一部分,该外显子长0.6×10³个碱基对。在最后一个外显子中有两个用于RNA剪接的典型受体位点,第一个位于外显子的5'端,第二个位于该端下游370个碱基对处。结合S1核酸酶图谱分析,上述结果使我们得出结论:通过这两个受体位点的差异使用,单个基因产生了两种不同的mRNA,分别为1.45×10³个碱基对和1.1×10³个碱基对,其中一种含有最后一个外显子的5'半部分,另一种则没有。这两种mRNA似乎利用相同的修饰聚腺苷酸化信号AGTAAA,而不是聚腺苷酸化位点下游约30个碱基对处存在的真实AATAAA序列。这可能是因为在距AGTAAA序列适当距离处存在另一个富含G + T的共有序列,而距AATAAA序列则没有。心肌肌球蛋白碱性轻链基因已被证明在心室肌、心房肌和背阔肌前部中表达,其中背阔肌前部是慢肌骨骼肌的特征。在这些肌肉中,表达了两种与心室肌中相同的心肌肌球蛋白碱性轻链mRNA,并且它们在每个组织中的相对含量与心室肌中的几乎相同。

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