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首页> 外文期刊>Journal of Cellular Physiology >Tyrosine Residues Regulate Multiple Nuclear Functions of P54nrb
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Tyrosine Residues Regulate Multiple Nuclear Functions of P54nrb

机译：酪氨酸残基调节多个核P54nrb功能

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摘要

The non-POU-domain-containing octamer binding protein (NONO; also known as p54nrb) has various nuclear functions ranging from transcription, RNA splicing, DNA synthesis and repair. Although tyrosine phosphorylation has been proposed to account for the multi-functional properties of p54nrb, direct evidence on p54nrb as a phosphotyrosine protein remains unclear. To investigate the tyrosine phosphorylation status of p54nrb, we performed site-directed mutagenesis on the five tyrosine residues of p54nrb, replacing the tyrosine residues with phenylalanine or alanine, and immunoblotted for tyrosine phosphorylation. We then preceded with luciferase reporter assays, RNA splicing minigene assays, co-immunoprecipitation, and confocal microscopy to study the function of p54nrb tyrosine residues on transcription, RNA splicing, protein-protein interaction, and cellular localization. We found that p54nrb was not phosphorylated at tyrosine residues. Rather, it has non-specific binding affinity to anti-phosphotyrosine antibodies. However, replacement of tyrosine with phenylalanine altered p54nrb activities in transcription co-repression and RNA splicing in gene context-dependent fashions by means of differential regulation of p54nrb protein association with its interacting partners and co-regulators of transcription and splicing. These results demonstrate that tyrosine residues, regardless of phosphorylation status, are important for p54nrb function. J. Cell. Physiol. 232: 852-861, 2017. (c) 2016 Wiley Periodicals, Inc.
机译：的non-POU-domain-containing八聚物结合蛋白质(禁忌;核函数,从转录RNA拼接、DNA合成和修复。提出了酪氨酸磷酸化占的多功能特性p54nrb p54nrb作为直接证据phosphotyrosine蛋白质仍不清楚。探讨酪氨酸磷酸化状态p54nrb,我们执行定点诱变p54nrb五个酪氨酸残基,取代的酪氨酸残基苯丙氨酸和丙氨酸,免疫印迹酪氨酸磷酸化。荧光素酶记者化验,RNA拼接小基因化验、co-immunoprecipitation和共焦显微镜研究p54nrb的功能酪氨酸残基上转录RNA拼接,蛋白质相互作用,和细胞本地化。在酪氨酸残基磷酸化。有非特异性亲和力anti-phosphotyrosine抗体。更换酪氨酸和苯丙氨酸在转录改变p54nrb活动co-repression和RNA剪接基因上下文相关的时尚的p54nrb微分调节蛋白质协会和它的合作伙伴和互动co-regulators转录、剪接。这些结果表明,酪氨酸残基,不管磷酸化状态对p54nrb功能很重要。232: 852 - 861, 2017。公司。

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