The proliferation potential protein-related (P2P-R) gene with domains encoding heterogeneous nuclear ribonucleoprotein association and Rb1 binding shows repressed expression during terminal differentiation.
Terminal differentiation is associated with repression in the expression of the proliferation potential proteins (P2P) subset of heterogeneous nuclear ribonucleoprotein (hnRNP) proteins. We report here the cloning and characterization of a 5173-bp P2P-related (P2P-R) cDNA that contains a 4214-bp open reading frame. Probes to this cDNA detect a single 8-kb mRNA in multiple murine tissues and in proliferating 3T3T cells, but not in terminally differentiated 3T3T adipocytes. Evidence that this cDNA can encode peptides with domains for hnRNP association was established by showing that such peptides are recognized by two monoclonal antibodies known to detect core hnRNP proteins, and by showing that the C130 monoclonal antibody, produced against a cDNA-derived fusion protein, also selectively detects native P2P hnRNP proteins. In addition, P2P-R cDNA-derived fusion proteins bind single-stranded nucleic acids, and a P2P-R cDNA-derived antisense oligonucleotide selectively represses P2P expression. Because terminal differentiation is associated with modulation in Rb1 function, we assayed if products of this cDNA might interact with Rb1. Evidence that the P2P-R cDNA encodes a protein domain that binds Rb1 was established using a glutathione S-transferase fusion protein to selectively precipitate Rb1 from cellular extracts. Data also show that this binding is reduced by competition with the adenovirus E1a protein, indicating that binding occurs through the "pocket" domain of Rb1. These results establish that the P2P-R cDNA encodes protein domains involved in both hnRNP association and Rb1 binding and complement recent reports that localize Rb1 to sites of RNA processing in the nucleus.
Witte
M M
MM
Department of Pathology, University of Tennessee Medical Center, Memphis 38163, USA.
Scott
R E
RE
D017667:Adipocytes / Q000166:cytology; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins; D002454:Cell Differentiation; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D005796:Genes; D006412:Hematopoietic Stem Cells / Q000166:cytology; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D016601:RNA-Binding Proteins; D016160:Retinoblastoma Protein / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D014018:Tissue Distribution
D002352:Carrier Proteins; D018076:DNA, Complementary; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C104647:PRKRA protein, human; D016601:RNA-Binding Proteins; C491290:Rbbp6 protein, mouse; D016160:Retinoblastoma Protein; D012261:Ribonucleoproteins