Molecular Cloning of cDNA for the Nuclear Ribonucleoprotein Particle C Proteins: A Conserved Gene Family

Abstract

The C proteins, C1 and C2, are major constituents of the heterogeneous nuclear RNA (hnRNA) ribonucleoprotein (hnRNP) complex in vertebrates. C1 and C2 are antigenically related phosphoproteins that are in contact with hnRNA in intact cells and bind to RNA tightly in vitro. A cDNA clone for the C proteins was isolated by immunological screening of a human lambda gt11 expression vector cDNA library with monoclonal antibodies. The lacZ-cDNA fusion protein is recognized by two different anti-C protein monoclonal antibodies. HeLa cell mRNA that was hybrid-selected with the cDNA clone (1.1 kilobases long) was translated in vitro and yielded both the C1 and C2 proteins (41 and 43 kDa, respectively). RNA blot analysis showed strong hybridization to two polyadenylylated transcripts, of about 1.4 kb and 1.9 kb, in human cells. Genomic DNA blot analysis showed multiple hybridizing restriction fragments in human and mouse, and homologous DNA sequences are found across eukaryotes from human to yeast. These findings suggest that the sequences encoding the hnRNP C proteins are members of a conserved gene family and they open the way for detailed molecular and genetic studies of these proteins.