Harvey ras genes transform without mutant codons, apparently activated by truncation of a 5' exon (exon -1).
The hypothesis is tested that the ras gene of Harvey sarcoma virus (Ha-SV) and the proto-ras DNAs from certain tumor cells derive transforming function from specific codons in which they differ from normal proto-ras genes. Molecularly cloned Harvey proviral vectors carrying viral ras, normal rat proto-ras, and recombinant ras genes in which the virus-specific ras codons 12 and 59 were replaced by proto-ras equivalents each transformed aneuploid mouse 3T3 cells after latent periods that ranged from 4 to 10 days. Viruses with or without virus-specific ras codons all transformed diploid rat cells in 3-5 days equally well. However, in the absence of virus replication, mutant codons were beneficial for transforming function. Deletion of non-ras regions of Ha-SV did not affect transforming function. We conclude that specific ras codons are not necessary for transforming function. Comparisons of the ras sequences of Ha-SV, BALB SV, and Rasheed SV with sequences of proto-ras genes from rat and man revealed an upstream proto-ras exon, termed exon -1. The 3' end of this exon is present in all three viruses and in a ras pseudogene of the rat. Since ras genes transform without mutation and since exon -1 is truncated in viral ras genes and all transforming proto-ras DNAs of the Harvey and the Kirsten ras family, we propose that ras genes are activated by truncation of exon -1 either via viral transduction or artificially via cloning and transfection. The proposal implies that untruncated proto-ras genes with point mutations may not be cellular cancer genes.
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D005786:Gene Expression Regulation; D005796:Genes; D051379:Mice; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D009857:Oncogenes; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D011519:Proto-Oncogenes
D003062:Codon; D009856:Oncogene Proteins, Viral; D011518:Proto-Oncogene Proteins