Isolation and characterization of a new 110 kDa human nuclear RNA-binding protein (p110nrb).
RNA-protein interactions play key roles in many fundamental cellular processes such as RNA processing, RNA transport, and RNA translation. During our attempts to isolate the human U6 small nuclear RNA capping enzyme, we identified a new 110 kDa nuclear RNA-binding protein, designated p110nrb. The full-length cDNA clone for p110nrb was characterized, and it encodes a 963 amino acid polypeptide. It is a highly acidic protein (pI 5.28) and the carboxyl terminal portion contains two conserved RNP motifs. A databank search found a putative C. elegans protein that might be the p110nrb homologue. The p110nrb was overexpressed as a glutathione S-transferase fusion protein in insect Sf9 cells, purified by affinity chromatography and injected into rabbits to produce specific polyclonal antibodies. Immunofluorescent staining showed that p110nrb is distributed evenly throughout the nucleoplasm. Northern blots showed that the mRNA is expressed in all tissues examined. An in vitro RNA-binding assay showed that p110nrb bound to RNA. These data suggest that p110nrb may play a role in the metabolism of nuclear RNA.
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA.
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J
Gu
S
S
Shimba
N
N
Nomura
R
R
Reddy
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm; D017173:Caenorhabditis elegans / Q000235:genetics; D002478:Cells, Cultured; D006801:Humans; D007313:Insecta / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification*; D015138:RNA, Nuclear / Q000737:chemistry / Q000302:isolation & purification*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D015138:RNA, Nuclear; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C416530:SART3 protein, human