THE HNRNP F-PROTEIN - UNIQUE PRIMARY STRUCTURE, NUCLEIC ACID-BINDING PROPERTIES, AND SUBCELLULAR-LOCALIZATION
THE HNRNP F-PROTEIN - UNIQUE PRIMARY STRUCTURE, NUCLEIC ACID-BINDING PROPERTIES, AND SUBCELLULAR-LOCALIZATION
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DOI：
10.1093/nar/22.6.1059
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发表时间：
1994-03-25
影响因子：
14.9
通讯作者：
DREYFUSS, G
中科院分区：
文献类型：
--
作者：
MATUNIS, MJ;XING, J;DREYFUSS, G
在哺乳动物细胞核中，有超过20种不同的异质核核糖核蛋白（hnRNP）与前体mRNA相关，这些蛋白质似乎影响前体mRNA的加工以及mRNA代谢和转运的其他方面。hnRNP蛋白在前mRNA上的排列对于每种RNA来说可能是独特的，并且可能由这些蛋白质中每种蛋白质的不同RNA结合偏好决定。hnRNP F（M（r）= 53 kD，pI = 6.1）和hnRNP H（M（r）= 56 kD，pI = 6.7-7.1）是免疫纯化的hnRNP复合物的丰富组分，它们具有不同的核酸结合特性。hnRNP F和hnRNP H在体外仅与poly（rG）结合，而其他hnRNP蛋白对不同的核糖核苷酸均聚物和ssDNA具有不同的亲和力。通过poly（ra）亲和层析从HeLa细胞中纯化hnRNP F和hnRNP H，并使用衍生自肽序列的寡核苷酸分离编码hnRNP F的cDNA。预测的hnRNP F的氨基酸序列揭示了一种新的蛋白质，具有与RNP共有序列RNA结合结构域相关的三个重复结构域。针对细菌表达的hnRNP F产生的单克隆抗体对hnRNP F和hnRNP H都是特异性的，并且在不同的生物体中识别相关蛋白，包括在酵母酿酒酵母中。因此，hnRNP F和hnRNP H在免疫学上高度相关，并且它们共享相同的肽。有趣的是，免疫荧光显微镜显示，hnRNP F和hnRNP H集中在离散区域的核质，在一般的核质分布以前的特点hnRNP蛋白。hnRNP F和hnRNP H独特的RNA结合特性、氨基酸序列和独特的核内定位使它们成为新的hnRNP蛋白，可能对含有富含鸟苷序列的RNA的加工是重要的。
More than 20 different heterogeneous nuclear ribonucleoproteins (hnRNPs) are associated with pre-mRNAs in the nucleus of mammalian cells and these proteins appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. The arrangement of hnRNP proteins on pre-mRNAs is likely to be unique for each RNA and may be determined by the different RNA-binding preferences of each of these proteins. hnRNP F (M(r) = 53 kD, pl = 6.1) and hnRNP H (M(r) = 56 kD, pl = 6.7-7.1) are abundant components of immunopurified hnRNP complexes and they have distinct nucleic acid binding properties. Unlike other hnRNP proteins which display a varying range of affinities for different ribonucleotidehomopolymers and ssDNA, hnRNP F and hnRNP H bind only to poly(rG) in vitro. hnRNP F and hnRNP H were purified from HeLa cells by poly(ra) affinity chromatography and oligonucleotides derived from peptide sequences were used to isolate a cDNA encoding hnRNP F. The predicted amino acid sequence of hnRNP F revealed a novel protein with three repeated domains related to the RNP consensus sequence RNA-binding domain. Monoclonal antibodies produced against bacterially expressed hnRNP F were specific for both hnRNP F and hnRNP H and recognized related proteins in divergent organisms, including in the yeast Saccharomyces cerevisiae. hnRNP F and hnRNP H are thus highly related immunologically and they share identical peptides. Interestingly, immunofluorescence microscopy revealed that hnRNP F and hnRNP H are concentrated in discrete regions of the nucleoplasm, in contrast to the general nucleoplasmic distribution of previously characterized hnRNP proteins. The unique RNA-binding properties, amino acid sequence and distinct intranuclear localization of hnRNP F and hnRNP H make them novel hnRNP proteins that are likely to be important for the processing of RNAs containing guanosine-rich sequences.