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首页> 外文期刊>Molecular and Cellular Biology >Transcription of a variant human U6 small nuclear RNA gene is controlled by a novel, internal RNA polymerase III promoter.
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Transcription of a variant human U6 small nuclear RNA gene is controlled by a novel, internal RNA polymerase III promoter.

机译：变异的人类U6小核RNA基因的转录受新型内部RNA聚合酶III启动子控制。

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Promoter elements in the 5' flanking regions of vertebrate U6 RNA genes have been shown to be both necessary and sufficient for transcription by RNA polymerase III. We have recently isolated and characterized a variant human U6 gene (87U6) that can be transcribed by RNA polymerase III in vitro in the absence of any natural 5' or 3' flanking sequences. This gene contains 10 nucleotide differences from the previously characterized human U6 gene (wtU6) within the coding region but has no homology to wtU6 in the upstream promoter region. By constructing chimeras between these two genes, we have shown that mutation of as few as two nucleotides in the coding region of the human U6 RNA gene is sufficient to create an internal promoter that is functional in vitro. A T-to-C transition at position 57 and a single T deletion at position 52 produce an internal U6 promoter that is nearly as active in vitro as the external U6 polymerase III promoter utilized by wtU6. Neither of these residues is absolutely conserved during evolution, and both of these nucleotide changes occur within the previously noted A box homology. Deletion and linker scanning mutations within the coding region of this variant U6 gene suggest that, in addition to the central region including bp 52 and 57, sequences at the extreme 5' end of the gene are critical for efficient transcription. In contrast, flanking sequences have a minor effect on transcriptional efficiency. This arrangement is unique among internal RNA polymerase III promoters and may indicate unique regulation of this gene.
机译：已显示脊椎动物U6 RNA基因5'侧翼区域的启动子元件对于RNA聚合酶III的转录既必要又足够。我们最近已经分离并鉴定了人类U6变异基因（87U6），可以在不存在任何天然5'或3'侧翼序列的情况下在体外通过RNA聚合酶III进行转录。该基因在编码区域内与先前表征的人U6基因（wtU6）包含10个核苷酸差异，但在上游启动子区域与wtU6没有同源性。通过在这两个基因之间构建嵌合体，我们已经表明，人U6 RNA基因编码区中少至两个核苷酸的突变就足以创建一个在体外具有功能的内部启动子。第57位的T-C转换和第52位的T缺失会产生一个内部U6启动子，其在体外的活性几乎与wtU6所使用的外部U6聚合酶III启动子相同。在进化过程中，这些残基都不是绝对保守的，并且这两个核苷酸变化均发生在先前提到的A box同源性内。该变体U6基因的编码区内的缺失和接头扫描突变表明，除了包括bp 52和57的中央区域之外，该基因的5'末端的序列对于有效转录至关重要。相反，侧翼序列对转录效率影响较小。这种排列在内部RNA聚合酶III启动子中是独特的，可能表明该基因的独特调控。

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