cDNA cloning and analysis of the expression of nucleoporin p45

The p62 complex is an assembly of four O-linked glycoproteins (p62, p58, p54, and p45) localized in the central region of the nuclear pore complex. It has been suggested to provide a substrate binding site near the central gated channel of the pore during nuclear protein import. The sequences of p62, p58, and p54 from rat have been reported previously. We have now carried out cDNA cloning of rat p45. The authenticity of the p45 clone was confirmed by two-dimensional gel analysis of the in vitro translated product of this clone. Sequence comparison showed that p45 is mostly identical to the amino terminal four-fifths of p58. p45 contains an N-terminal FG (Phe-Gly) repeat region, a middle coiled-coil region, and a truncated C-terminal FG repeat region (compared to p58). The sequence data and genomic Southern hybridization results strongly support the possibility that p45 and p58 are generated by mRNA alternative splicing. The sequences of three other p58-related cDNA clones indicate that the p58/p45 gene transcript gives rise to additional alternatively spliced mRNAs in mammalian cells. Interestingly, the expression level of p45 relative to p58 varies in different cultured cell lines, indicating that the p62 complex is heterogeneous with respect to these two subunits.