RNA-binding protein RBM8A (Y14) and MAGOH localize to centrosome in human A549 cells
RNA-binding protein RBM8A (Y14) and MAGOH localize to centrosome in human A549 cells
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DOI：
10.1007/s00418-013-1135-4
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发表时间：
2014-01-01
影响因子：
2.3
通讯作者：
Tomosugi, Naohisa
中科院分区：
文献类型：
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作者：
Ishigaki, Yasuhito;Nakamura, Yuka;Tomosugi, Naohisa
RBM 8A（Y14）携带RNA结合基序，与MAGOH形成紧密的异源二聚体。已知异二聚体是输出mRNA上的外显子连接复合物的成员，并且是mRNA代谢如剪接、mRNA输出和无义介导的mRNA降解所需的。几乎所有RBM 8A-MAGOH复合物都定位于核质中，并在细胞核和细胞质之间穿梭以进行RNA代谢。最近，有报道称RBM 8A或MAGOH缺陷细胞中存在G2/M转换异常和异常中心体调节。这些结果促使我们重新评估RBM 8A-MAGOH在人细胞中的定位。有趣的是，我们的免疫染色实验表明，这些蛋白质的定位在中心体除了核。此外，瞬时表达的eYFP标记的RBM 8A和Flag标记的MAGOH也与中心体信号共定位。此外，通过原位邻位连接法检测中心体复合物的形成。我们的实验清楚地表明，Myc标记的RBM 8A和Flag标记的MAGOH在中心体中形成复合物。GFP标记的PLK 1也与Myc-RBM 8A共定位。我们的研究结果表明，RBM 8A-MAGOH复合物是通过直接定位到中心体而不是间接影响M期进展所必需的。
RBM8A (Y14) is carrying RNA-binding motif and forms the tight heterodimer with MAGOH. The heterodimer is known to be a member of exon junction complex on exporting mRNA and is required for mRNA metabolisms such as splicing, mRNA export and nonsense-mediated mRNA decay. Almost all RBM8A-MAGOH complexes localize in nucleoplasm and shuttle between nuclei and cytoplasm for RNA metabolism. Recently, the abnormality of G2/M transition and aberrant centrosome regulation in RBM8A- or MAGOH-deficient cells has been reported. These results prompt us to the reevaluation of the localization of RBM8A-MAGOH in human cells. Interestingly, our immunostaining experiments showed the localization of these proteins in centrosome in addition to nuclei. Furthermore, the transiently expressed eYFP-tagged RBM8A and Flag-tagged MAGOH also co-localized with centrosome signals. In addition, the proximity ligation in situ assay was performed to detect the complex formation in centrosome. Our experiments clearly showed that Myc-tagged RBM8A and Flag-tagged MAGOH formed a complex in centrosome. GFP-tagged PLK1 also co-localized with Myc-RBM8A. Our results show that RBM8A-MAGOH complex is required for M-phase progression via direct localization to centrosome rather than indirect effect.