Identification of the COMM-domain containing protein 1 as specific binding partner for the guanine-rich RNA sequence binding factor 1
Identification of the COMM-domain containing protein 1 as specific binding partner for the guanine-rich RNA sequence binding factor 1
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DOI：
10.1016/j.bbagen.2020.129678
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发表时间：
2020-11-01
影响因子：
3
通讯作者：
Sofi, Sajad
中科院分区：
文献类型：
--
作者：
Dumoulin, Bernhard;Ufer, Christoph;Sofi, Sajad
工作背景：富含鸟嘌呤的RNA序列结合因子1（GRSF 1）是hnRNP H/F家族的RNA结合蛋白，其已经涉及红细胞生成、氧化还原稳态的调节、胚胎脑发育、线粒体功能和细胞衰老。GRSF 1-RNA相互作用的分子基础在过去已经被广泛研究，但目前GRSF 1结合蛋白尚未被鉴定。方法：为了寻找GRSF 1结合蛋白，我们首先利用酵母双杂交系统，从人胎脑cDNA文库中筛选潜在的GRSF 1结合蛋白。随后，我们探索了重组蛋白的蛋白质-蛋白质相互作用，进行了免疫沉淀实验以证实活细胞中天然蛋白质的相互作用，并进行了截短研究以鉴定GRSF 1的蛋白质结合基序。利用酵母双杂交系统，我们鉴定了COMMD 1蛋白。作为特异性GRSF 1结合蛋白，体外截短研究表明COMMD 1与GRSF 1富含丙氨酸的结构域相互作用。结论：在哺乳动物细胞中，COMMD 1与GRSF 1的富含丙氨酸的结构域以RNA非依赖的方式特异性相互作用。一般意义：这是第一份描述特异性GRSF 1结合蛋白的报告。一般意义：这是第一份描述特异性GRSF 1结合蛋白的报告。
Background: The guanine-rich RNA sequence binding factor 1 (GRSF1) is an RNA-binding protein of the hnRNP H/F family, which has been implicated in erythropoiesis, regulation of the redox homeostasis, embryonic brain development, mitochondrial function and cellular senescence. The molecular basis for GRSF1-RNA interaction has extensively been studied in the past but for the time being GRSF1 binding proteins have not been identified.Methods: To search for GRSF1 binding proteins we first employed the yeast two-hybrid system and screened a cDNA library of human fetal brain for potential GRSF1 binding proteins. Subsequently, we explored the protein-protein-interaction of the recombiant proteins, carried out immunoprecipitation experiments to confirm the interaction of the native proteins in living cells and performed truncation studies to identify the protein-binding motif of GRSF1.Results: Using the yeast two-hybrid system we identified the COMM-domain containing protein 1 (COMMD1) as specific GRSF1 binding protein and in vitro truncation studies suggested that COMMD1 interacts with the alanine-rich domain of GRSF1. Co-immunoprecipitation strategies indicated that COMMD1-GRSF1 interaction was RNA independent and also occurred in living cells expressing the two native proteins.Conclusion: In mammalian cells the COMM-domain containing protein 1 (COMMD1) specifically interacts with the Ala-rich domain of GRSF1 in an RNA-independent manner. General significance: This is the first report describing a specific GRSF1 binding protein.General significance: This is the first report describing a specific GRSF1 binding protein.