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首页> 外文期刊>Molecular and Cellular Biology >Different forms of Go alpha mRNA arise by alternative splicing of transcripts from a single gene on human chromosome 16.
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Different forms of Go alpha mRNA arise by alternative splicing of transcripts from a single gene on human chromosome 16.

机译：Go alpha mRNA的不同形式是通过人类16号染色体上单个基因的转录产物的可变剪接产生的。

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摘要

Go alpha, (gene symbol GNA01), a member of the signal-transducing guanine nucleotide-binding (G) protein family, has been implicated in ion channel regulation. Some tissues contain multiple Go alpha mRNAs of different sizes that differ in the 3' untranslated regions (UTRs). Using sequence-specific 48-base oligonucleotides, two complementary to the different 3' UTRs and one complementary to the coding region, we investigated the origin of the multiple Go alpha transcripts, the organization of the Go alpha gene, the interspecies conservation of 3' UTRs, and the chromosomal localization of Go alpha. Oligonucleotides labeled to high specific activity by using terminal deoxynucleotidyltransferase each hybridized with a single band of restriction enzyme-digested mouse and human DNAs. In three of four digests of human DNA, the two probes specific for the different 3' UTRs hybridized with the same restriction fragment. Thus, these nucleotide sequences are in close proximity in the human genome. The order of the UTRs in the bovine, human, and mouse genomes was confirmed directly by polymerase chain reaction (PCR) amplification and sequencing. Hybridization of bovine oligonucleotide sequence with mouse and human genomic DNA indicated a high degree of interspecies sequence conservation: conservation was confirmed by PCR amplification and sequencing. Bands detected by both UTR probes, as well as the predominant bands detected by a bovine Go alpha cDNA, segregated with human chromosome 16 on Southern blot analysis of human-mouse somatic cell hybrids. We conclude that Go alpha mRNAs with different 3' UTRs arise by alternative splicing of transcripts from a single gene. The UTRs, which exhibit a high degree of interspecies conservation, may play a role in regulation of Go alpha expression during differentiation or in specific tissues. The use of oligonucleotide probes of the type described here represents a new strategy, potentially widely applicable for mapping and elucidating structural features of genes.
机译：Go alpha（基因符号GNA01）是信号传导鸟嘌呤核苷酸结合（G）蛋白家族的成员，已参与离子通道调控。一些组织包含3'非翻译区（UTR）不同的多个大小不同的Go alpha mRNA。使用序列特异性的48个碱基的寡核苷酸，两个互补于不同的3'UTR，一个互补于编码区，我们研究了多个Go alpha转录本的起源，Go alpha基因的组织，3'的种间保守性UTR和Go alpha的染色体定位。通过使用末端脱氧核苷酸转移酶将寡核苷酸标记为高比活性，每个末端寡核苷酸都与单条限制性酶消化的小鼠和人类DNA杂交。在人类DNA的四个消化物中的三个中，对不同3'UTR特异的两个探针与相同的限制性片段杂交。因此，这些核苷酸序列在人类基因组中非常接近。牛，人和小鼠基因组中UTR的顺序直接通过聚合酶链反应（PCR）扩增和测序确定。牛寡核苷酸序列与小鼠和人类基因组DNA的杂交表明种间序列的高度保守：通过PCR扩增和测序证实了保守。在人-小鼠体细胞杂交体的Southern印迹分析中，两种UTR探针检测到的条带以及牛Go alpha cDNA检测到的主要条带与16号人类染色体分离。我们得出的结论是，具有不同3'UTR的Go alpha mRNA通过来自单个基因的转录物的可变剪接产生。具有高度种间保守性的UTRs可能在分化过程中或特定组织中调控Go alpha表达。本文所述类型的寡核苷酸探针的使用代表了一种新的策略，可能广泛适用于作图和阐明基因的结构特征。

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