A heteroduplex study of the sequence relationships between the RNAs of M-MSV and M-MLV

Abstract

The regions of sequence homology between the RNA genomes of a murine sarcoma virus (clone 124 Moloney-MSV) and its parental helper virus (clone 3 Moloney-murine leukemia virus (M-MLV)) have been mapped. Long complementary DNA transcripts of the MMLV RNA were hybridized to M-MSV RNA, and the structures of the hybrids were observed in the electron microscope. Beginning at the 5′ end, the two RNAs are homologous for a region of length 2.25 kb (kilobases). In the next region, of length approximately 4.2 kb on the MLV genome, there are several homology segments between MLV and MSV, but there are also several short sequences present on MLV and deleted in MSV. There is then a major substitution loop, with a sequence (β L) of length 2.9 kb present on MLV and missing on MSV, and a sequence (β S) of length 1.5 kb present on MSV and missing on MLV. At the 3′ end, there is a homology sequence of length 0.8 kb. On the basis of these results, other data on genes expressed in M-MSV-transformed cells, and by analogy with the avian gene map, we suggest that the gag genes (internal structural proteins) lie in the 2.25 kb region of homology near the 5′ ends of M-MSV and M-MLV RNAs, and that the β S segment contains a sarcoma ( src) gene. Some of the heteroduplexes and some of the MLV cDNA/MLV RNA homoduplexes are circular, thus showing that cDNA transcription is initiated at an internal position in the RNA, proceeds to the 5′ end, and then “jumps” to the 3′ end.