Identification of the RNA recognition element of the RBPMS family of RNA-binding proteins and their transcriptome-wide mRNA targets
Identification of the RNA recognition element of the RBPMS family of RNA-binding proteins and their transcriptome-wide mRNA targets
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DOI：
10.1261/rna.045005.114
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发表时间：
2014-07-01
期刊：
影响因子：
4.5
通讯作者：
Tuschl, Thomas
中科院分区：
文献类型：
--
作者：
Farazi, Thalia A.;Leonhardt, Carl S.;Tuschl, Thomas
最近的研究表明，rna结合蛋白与多重剪接（RBPMS）家族蛋白在卵母细胞、视网膜神经节细胞、心脏和胃肠道平滑肌发育中起着重要作用。这些RNA结合蛋白含有单个RNA识别基序（RRM），其靶点和分子功能尚未被确定。我们在HEK293细胞中使用光激活核糖核苷增强交联和免疫沉淀（PAR-CLIP）定义了转录组范围内的RNA靶点，揭示了外显子成熟和内含子前mrna结合位点，与蛋白质的核和细胞质定位一致。计算和生化方法将RNA识别元件（RRE）定义为由可变间隔区分隔的串联CAC三核苷酸基序。与其他mRNA结合蛋白类似，RBPMS蛋白家族在氧化应激条件下重新定位到细胞质应激颗粒，这表明它在大核和/或多核细胞中具有支持mRNA定位的功能，并优先表达。
Recent studies implicated the RNA-binding protein with multiple splicing (RBPMS) family of proteins in oocyte, retinal ganglion cell, heart, and gastrointestinal smooth muscle development. These RNA-binding proteins contain a single RNA recognition motif (RRM), and their targets and molecular function have not yet been identified. We defined transcriptome-wide RNA targets using photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) in HEK293 cells, revealing exonic mature and intronic pre-mRNA binding sites, in agreement with the nuclear and cytoplasmic localization of the proteins. Computational and biochemical approaches defined the RNA recognition element (RRE) as a tandem CAC trinucleotide motif separated by a variable spacer region. Similar to other mRNA-binding proteins, RBPMS family of proteins relocalized to cytoplasmic stress granules under oxidative stress conditions suggestive of a support function for mRNA localization in large and/or multinucleated cells where it is preferentially expressed.