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非洲爪蟾U7小核RNA编码基因具有异常紧密的结构。

The Xenopus U7 snRNA-encoding gene has an unusually compact structure.

作者信息

Watkins N J, Phillips S C, Turner P C

机构信息

Department of Biochemistry, University of Liverpool, UK.

出版信息

Gene. 1992 Oct 21;120(2):271-6. doi: 10.1016/0378-1119(92)90104-w.

Abstract

A subclone containing a single Xenopus borealis U7 snRNA-encoding gene has been microinjected into X. laevis oocyte nuclei to examine its expression using [32P]GTP as an in vivo label. Only two U7 snRNA bands were detected after incubation, and subsequent fractionation of the oocyte showed that only the larger transcript is present in the nucleus. The sequence of this functional U7 gene shows that, in addition to the coding region, it contains, in the appropriate locations, the 3'-box and proximal sequence element (PSE) which are typical of Pol II-transcribed snRNA genes. Surprisingly, the Xenopus U7 gene contains two adjacent octamer-binding motifs located only 12 and 24 bp upstream from the PSE, instead of the usual location around 150-200 bp upstream. No other cis-acting elements appear to be present. A 5' deletion analysis shows that the transcription level of this U7 gene remains constant if sequences upstream of the two octamer motifs are removed, yet is undetectable when an additional 34 bp containing both octamers and the PSE are removed. This confirms that the Xenopus U7 gene is the most compact snRNA-encoding gene isolated to date. A comparison of U7 sequences shows there is a much greater conservation in the 5' half of the molecule, which contains sequences that base-pair with target pre-mRNA, than in the 3' half which can form a single stem-loop structure that varies in size.

摘要

一个含有单个非洲爪蟾U7小核仁RNA编码基因的亚克隆已被显微注射到非洲爪蟾卵母细胞核中,以使用[32P]GTP作为体内标记来检测其表达。孵育后仅检测到两条U7小核仁RNA条带,随后对卵母细胞进行分级分离显示,只有较大的转录本存在于细胞核中。这个功能性U7基因的序列表明,除了编码区外,它在适当位置还包含3'-框和近端序列元件(PSE),这是典型的由RNA聚合酶II转录的小核仁RNA基因。令人惊讶的是,非洲爪蟾U7基因在PSE上游仅12和24 bp处含有两个相邻的八聚体结合基序,而不是通常位于上游约150 - 200 bp处的位置。似乎不存在其他顺式作用元件。5'缺失分析表明,如果去除两个八聚体基序上游的序列,该U7基因的转录水平保持不变,但当去除包含两个八聚体和PSE的另外34 bp时则检测不到。这证实了非洲爪蟾U7基因是迄今为止分离出的最紧凑的小核仁RNA编码基因。U7序列的比较表明,分子的5'半部分(包含与靶前体mRNA碱基配对的序列)比3'半部分(可形成大小不同的单个茎环结构)具有更高的保守性。

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