Nuclear Pnn/DRS protein binds to spliced mRNPs and participates in mRNA processing and export via interaction with RNPS1.
Pnn/DRS protein is associated with desmosomes and colocalizes with splicing factors in nuclear speckled domains. The potential interaction of Pnn with RNPS1, a pre-mRNA splicing factor and a component of the exon-exon junction complex, prompted us to examine whether Pnn is involved in nuclear mRNA processing. By immunoprecipitation, we found that Pnn associates preferentially with mRNAs produced by splicing in vitro. Oligonucleotide-directed RNase H digestion revealed that Pnn binds to the spliced mRNAs at a position immediately upstream of the splice junction and that 5' splice site utilization determines the location of Pnn in alternatively spliced mRNAs. Immunoprecipitation further showed that Pnn binds to mRNAs produced from a transiently expressed reporter in vivo. Although associated with mRNPs, Pnn is a nuclear-restricted protein as revealed by the heterokaryon assay. Overexpression of an amino-terminal fragment of Pnn that directly interacts with RNPS1 leads to blockage of pre-mRNA splicing. However, although suppression of Pnn expression shows no significant effect on splicing, it leads to some extent to nuclear accumulation of bulk poly(A)(+) RNA. Therefore, Pnn may participate, via its interaction with RNPS1, in mRNA metabolism in the nucleus, including mRNA splicing and export.
D001692:Biological Transport; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007438:Introns; D008954:Models, Biological; D009687:Nuclear Proteins / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D014162:Transfection
D015815:Cell Adhesion Molecules; D009687:Nuclear Proteins; C102646:PNN protein, human; D012333:RNA, Messenger; C084538:RNPS1 protein, human; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D016914:Ribonuclease H