Nucleotide sequence of the first exon of the rat c-myc gene: proviral insertions in murine leukemia virus-induced lymphomas do not affect exon 1.

We have previously reported that proviruses are integrated adjacent to the c-myc gene in rat thymomas induced by murine leukemia viruses. In order to characterize these insertions, we have isolated recombinant DNA clones from normal rat DNA containing all of the normal rat c-myc gene, and from two Moloney murine leukemia virus-induced lymphomas containing both proviral and adjacent rat c-myc sequences. We determined the DNA sequence of portions of the normal and one tumor-derived clone. The normal and tumor-derived exon 1 sequences are identical. By comparing our sequence to the sequences of mouse and human c-myc, we located the first exon of the rat c-myc gene. Analysis of the tumor-derived rat c-myc clones showed that proviral integration occurred approximately 1.4 kb upstream of exon 1 of c-myc in the case of one tumor and 0.55 kb upstream of c-myc exon 1 in the other. Thus, we conclude that the proviral insertions in these tumors did not affect the rat c-myc gene by altering the structure of the c-myc RNA. Consistent with this, the c-myc RNA present in a cell line derived from one of these tumors is identical in size to the normal c-myc RNA. Furthermore, the level of c-myc expression is not dramatically elevated in this cell line. Exon 1 of the rat c-myc gene contains no ATG start codons and contains multiple stop codons in all three reading frames, indicating that it, like the chicken and mouse exon 1 sequences, is noncoding. The extent of homology between our sequence of rat c-myc exon 1 and the published sequence of human c-myc exon 1 is similar to the extent of homology between the sequences of mouse and human c-myc exon 1. The rat and mouse c-myc exon 1 sequences differ from each other by about the amount predicted from the known divergence times of mice from rats. Exon 1 of c-myc is only slightly conserved, evolving at a rate similar to that seen for introns and pseudogenes.