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大鼠肾脏带3型氯离子/碳酸氢根离子交换体信使核糖核酸由一个可变启动子转录而来。

Rat kidney band 3 Cl-/HCO3- exchanger mRNA is transcribed from an alternative promoter.

作者信息

Kudrycki K E, Shull G E

机构信息

Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Ohio 45267-0524.

出版信息

Am J Physiol. 1993 Mar;264(3 Pt 2):F540-7. doi: 10.1152/ajprenal.1993.264.3.F540.

Abstract

We have previously shown that the rat kidney band 3 Cl-/HCO3- exchanger mRNA encodes an NH2-terminal truncated form of band 3 and that its 5' end differs from that of the erythrocyte band 3 mRNA (K. E. Kudrycki and G. E. Shull. J. Biol. Chem. 264: 8185-8192, 1989). To determine the genetic basis for the alternative kidney and erythroid mRNAs, we 1) isolated and characterized a rat erythroid band 3 cDNA, 2) isolated the rat band 3 gene and determined the exon/intron organization of sequences corresponding to the alternative 5' ends of the rat kidney and erythroid mRNAs, and 3) identified the transcription initiation sites of the two transcripts. The unique sequences at the 5' end of the rat erythroid mRNA are derived from exons 1-3 and are followed directly by sequences from exon 4 that are common to both mRNAs. In the kidney mRNA, sequences upstream of exon 4 are derived entirely from intron 3. Primer extension and S1 nuclease protection analyses demonstrate the presence of multiple transcription initiation sites for the rat erythroid band 3 mRNA at the beginning of exon 1, whereas the transcription initiation site for the kidney mRNA is located within intron 3. Thus two distinct promoters, separated by almost 5 kb of genomic sequence, are responsible for the highly tissue-specific transcription of the alternative rat erythroid and kidney band 3 mRNAs.

摘要

我们先前已经表明,大鼠肾脏带3 Cl⁻/HCO₃⁻交换体mRNA编码一种氨基末端截短形式的带3,并且其5'端与红细胞带3 mRNA的5'端不同(K.E. Kudrycki和G.E. Shull。《生物化学杂志》264: 8185 - 8192, 1989)。为了确定肾脏和红细胞mRNA差异的遗传基础,我们进行了以下操作:1)分离并鉴定了大鼠红细胞带3 cDNA;2)分离了大鼠带3基因,并确定了与大鼠肾脏和红细胞mRNA的可变5'端相对应的序列的外显子/内含子组织;3)确定了两种转录本的转录起始位点。大鼠红细胞mRNA 5'端的独特序列来自外显子1 - 3,紧接着是两种mRNA共有的外显子4的序列。在肾脏mRNA中,外显子4上游的序列完全来自内含子3。引物延伸和S1核酸酶保护分析表明,大鼠红细胞带3 mRNA在外显子1起始处存在多个转录起始位点,而肾脏mRNA的转录起始位点位于内含子3内。因此,被近5 kb基因组序列隔开的两个不同启动子,负责大鼠红细胞和肾脏带3 mRNA的高度组织特异性转录。

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