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首页> 外文期刊>Molecular and Cellular Biology >Characterization of SRp46, a Novel Human SR Splicing Factor Encoded by a PR264/SC35 Retropseudogene
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Characterization of SRp46, a Novel Human SR Splicing Factor Encoded by a PR264/SC35 Retropseudogene

机译：SRp46，由PR264 / SC35 Retropseudogene编码的新型人类SR剪接因子的表征。

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摘要

The highly conserved SR family contains a growing number of phosphoproteins acting as both essential and alternative splicing factors. In this study, we have cloned human genomic and cDNA sequences encoding a novel SR protein designated SRp46. Nucleotide sequence analyses have revealed that the SRp46 gene corresponds to an expressed PR264/SC35 retropseudogene. As a result of mutations and amplifications, the SRp46 protein significantly differs from the PR264/SC35 factor, mainly at the level of its RS domain. Northern and Western blot analyses have established that SRp46 sequences are expressed at different levels in several human cell lines and normal tissues, as well as in simian cells. In contrast, sequences homologous to SRp46 are not present in mice. In vitro splicing studies indicate that the human SRp46 recombinant protein functions as an essential splicing factor in complementing a HeLa cell S100 extract deficient in SR proteins. In addition, complementation analyses performed with β-globin or adenovirus E1A transcripts and different splicing-deficient extracts have revealed that SRp46 does not display the same activity as PR264/SC35. These results demonstrate, for the first time, that an SR splicing factor, which represents a novel member of the SR family, is encoded by a functional retropseudogene.
机译：高度保守的SR家族包含越来越多的磷蛋白，它们既是必需的剪接因子，又是替代剪接因子。在这项研究中，我们已经克隆了人类基因组和cDNA序列，这些序列编码一种称为SRp46的新型SR蛋白。核苷酸序列分析表明，SRp46基因对应于表达的PR264 / SC35 Retropseudogene。作为突变和扩增的结果，SRp46蛋白与PR264 / SC35因子存在显着差异，主要在其RS结构域水平上。 Northern和Western印迹分析已确定SRp46序列在几种人类细胞系和正常组织以及猿猴细胞中以不同的水平表达。相反，在小鼠中不存在与SRp46同源的序列。体外剪接研究表明，人SRp46重组蛋白在补充缺乏SR蛋白的HeLa细胞S100提取物中起着必要的剪接因子的作用。另外，用β-珠蛋白或腺病毒E1A转录本和不同的剪接缺陷提取物进行的互补分析显示，SRp46与PR264 / SC35的活性不同。这些结果首次证明，代表SR家族新成员的SR剪接因子由功能性逆转录假基因编码。

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