tailieunhanh - Báo cáo Y học: Importin a binds to an unusual bipartite nuclear localization signal in the heterogeneous ribonucleoprotein type I

The heterogeneous nuclear ribonucleoprotein (hnRNP) type I, a modulator of alternative splicing, localizes in the nucleoplasm of mammalian cells and in a discrete perinucleolar structure. HnRNP I contains a novel type of bipartite nuclear localization signal (NLS) at the N-terminus of the protein that we have previously named nuclear determinant localization type I (NLD-I). Recently, a neural counterpart of hnRNP I has been identified that contains a putative NLS with two strings of basic amino acids separated by a spacer of 30 residues | Eur. J. Biochem. 269 2727-2734 2002 FEBS 2002 doi Importin a binds to an unusual bipartite nuclear localization signal in the heterogeneous ribonucleoprotein type I Maria G. Romanelli and Carlo Morandi Department of Mother and Child Biology and Genetics University of Verona Italy The heterogeneous nuclear ribonucleoprotein hnRNP type I a modulator of alternative splicing localizes in the nucleoplasm of mammalian cells and in a discrete perinucleolar structure. HnRNP I contains a novel type of bipartite nuclear localization signal NLS at the N-terminus of the protein that we have previously named nuclear determinant localization type I NLD-I . Recently a neural counterpart of hnRNP I has been identified that contains a putative NLS with two strings of basic amino acids separated by a spacer of 30 residues. In the present study we show that the neural hnRNP I NLS is necessary and sufficient for nuclear localization and represents a variant of the novel bipartite NLS present in the NLD-I domain. Furthermore we demonstrate that the NLD-I is transported into the nucleus by cytoplasmic factor s with active transport modality. Binding assays using recombinant importin a show an interaction with NLD-I similar to that of SV40 large T antigen NLS. Deletion analysis indicates that both stretches of basic residues are necessary for binding to importin a. The above experimental results lead to the conclusion that importin a acts as cytoplasmic receptor for proteins characterized by a bipartite NLS signal that extends up to 37 residues. Keywords heterogeneous ribonucleoprotein-I polypyrimidine tract-binding protein PTB nuclear localization signal importin a. Transport of proteins and RNA into and out of the nucleus occurs through nuclear pore complexes NPCs which are plugged through the double membrane of the nuclear envelope 1 2 . Small molecules and ions may pass the NPC passively while macromolecules larger than 40-45 kDa are actively .

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