Characterization of ret proto-oncogene mRNAs encoding two isoforms of the protein product in a human neuroblastoma cell line.

The ret proto-oncogene expresses four major mRNA species of different lengths in human malignant cell lines and rat tissues. We isolated ret proto-oncogene cDNA clones from a cDNA library of a human neuroblastoma line, Nagai, which over-expressed these mRNAs. Four cDNA clones differing from each other in their 3' portions were analysed. The sequence of the region common to the cDNA clones is essentially identical to a reported cDNA sequence derived from THP-1 monocytic leukemia cells, that encodes a protein with characteristic features of receptor-type tyrosine kinase. From the 3' heterogeneity, two isoforms of the ret proto-oncogene product of 1072 and 1114 residues that differed from each other in their 9 and 51 C-terminal amino acids are predicted. Comparison of the structures of cDNA clones with that of the genomic clone showed that the 3' heterogeneity is produced by alternative polyadenylation and splicing of mRNA. Northern blot analysis using various fragments of cDNA indicated that the 4.5 kb, 3.9 kb and possibly 7.0 kb transcripts may encode a protein of 1072 residues, while the 6.0 kb transcript and a (minor) 4.6 kb transcript may encode a protein of 1114 residues.