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Abstract：
Segment 8 mRNAs of influenza virus A/Brevig Misson/1918/1 (H1N1) are poorly spliced compared to segment 8 mRNAs of influenza virus A/Netherlands/178/95 (H3N2). Using oligonucleotide-mediated protein pull down with oligos spanning the entire length of segment 8 of either influenza virus H1N1 or influenza virus H3N2 we identified cellular RNA binding proteins that interacted with oligonucleotides derived from either H1N1 or H3N2 sequences. When the identified hot spots for RNA binding proteins in H1N1 segment 8 mRNAs were replaced by H3N2 sequences, splicing efficiency increased significantly. Replacing as few as three nucleotides of the H1N1 mRNA with sequences from H3N2 mRNA, enhanced splicing of the H1N1 mRNAs. Cellular proteins U2AF65 and HuR interacted preferentially with the 3\'-splice site of H3N2 and overexpression of HuR reduced the levels of unspliced H1N1 mRNAs, suggesting that U2AF65 and HuR contribute to control of influenza virus mRNA splicing.
摘要：
与流感病毒A/荷兰/178/95(H3N2)的片段8mRNA相比,流感病毒A/BrevigMisson/1918/1(H1N1)的片段8mRNA的剪接较差。使用跨越流感病毒H1N1或流感病毒H3N2的片段8的整个长度的寡核苷酸介导的蛋白拉下,我们鉴定了与源自H1N1或H3N2序列的寡核苷酸相互作用的细胞RNA结合蛋白。当确定的热点RNA结合蛋白在H1N1片段8mRNA被H3N2序列取代，拼接效率显著提高。用H3N2mRNA的序列替换H1N1mRNA的仅三个核苷酸，H1N1mRNA的增强剪接。细胞蛋白U2AF65和HuR优先与H3N2的3'-剪接位点相互作用，HuR的过表达降低了未剪接的H1N1mRNA的水平，提示U2AF65和HuR有助于流感病毒mRNA剪接的控制。
