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Identification of novel ribonucleo-protein complexes from the brain-specific snoRNA MBII-52

机译:从大脑特异性snoRNA MBII-52鉴定新型核糖核蛋白复合物

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摘要

Small nucleolar RNAs (snoRNAs) guide nucleotide modifications within ribosomal RNAs or spliceosomal RNAs by base-pairing to complementary regions within their RNA targets. The brain-specific snoRNA MBII-52 lacks such a complementarity to rRNAs or snRNAs, but instead has been reported to target the serotonin receptor 2C pre-mRNA, thereby regulating pre-mRNA editing and/or alternative splicing. To understand how the MBII-52 snoRNA might be involved in these regulatory processes, we isolated the MBII-52 snoRNP from total mouse brain by an antisense RNA affinity purification approach. Surprisingly, by mass spectrometry we identified 17 novel candidates for MBII-52 snoRNA binding proteins, which previously had not been reported to be associated with canonical snoRNAs. Among these, Nucleolin and ELAVL1 proteins were confirmed to independently and directly interact with the MBII-52 snoRNA by coimmunoprecipitation. Our findings suggest that the MBII-52 snoRNA assembles into novel RNA-protein complexes, distinct from canonical snoRNPs.
机译:小核仁RNA(snoRNA)通过与RNA靶标内的互补区碱基配对,指导核糖体RNA或剪接体RNA内的核苷酸修饰。脑特异性snoRNA MBII-52缺乏与rRNA或snRNA的互补性,但是据报道,它靶向血清素受体2C pre-mRNA,从而调节pre-mRNA编辑和/或其他剪接。为了了解MBII-52 snoRNA可能如何参与这些调控过程,我们通过反义RNA亲和纯化方法从整个小鼠大脑中分离了MBII-52 snoRNP。出乎意料的是,通过质谱,我们鉴定了MBII-52 snoRNA结合蛋白的17种新型候选物,以前尚未报道其与规范性snoRNAs相关。其中,通过共免疫沉淀法确认了核仁蛋白和ELAVL1蛋白独立且直接与MBII-52 snoRNA相互作用。我们的发现表明,MBII-52 snoRNA组装成新颖的RNA-蛋白质复合物,与经典的snoRNP不同。

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