title: Morphf mutants of Rous sarcoma virus: nucleotide sequencing analysis suggests that a class of morphf mutants was generated through splicing of a cryptic intron.
abstract: The nature of the lesions involved in producing the fusiform phenotype of three mutants (WO101, WO201, and tsST529) of the Schmidt-Ruppin A strain of Rous sarcoma virus (RSV) was determined by molecular cloning and DNA sequencing. WO101 and WO201 contained an in-frame deletion of the v-src region coding for amino acids 116 to 140 of p60v-src. The deleted segment was flanked by consensus splice donor and acceptor sequences and contained an appropriately positioned branchpoint acceptor consensus sequence, suggesting that the deletion occurred through an aberrant RNA splicing event. S1 mapping experiments performed on RNA isolated from chicken cells infected with molecularly cloned wild-type RSV DNA suggested that the splice acceptor involved in the generation of this deletion was utilized at a low frequency (less than 1.0%) in wild-type RSV-infected cells. These results suggested that stable mutations may have arisen in the coding sequence of a eucaryotic viral transforming gene as a result of a probable aberrant RNA splicing event followed by reverse transcription into DNA. ST529 was found to harbor the same deletion present in WO101 and WO201 but also contained a point mutation which resulted in the substitution of lysine for glutamic acid at position 93. This change and the resulting large change in local charge were presumably required for the temperature-sensitive transformation phenotype of ST529. These results, together with other known deletions that produce fusiform mutants, suggested that a region within the amino-terminal one-third coding region of the src gene contributed to a structural domain of p60v-src that was important for controlling some morphological parameters of transformation in cells infected with RSV.
mesh_terms: D001358:Avian Sarcoma Viruses / Q000235:genetics*; D001483:Base Sequence; D002472:Cell Transformation, Viral; D002872:Chromosome Deletion; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D007438:Introns; D009154:Mutation; D009710:Nucleotide Mapping; D015688:Oncogene Protein pp60(v-src); D012326:RNA Splicing; D012367:RNA, Viral / Q000235:genetics; D012191:Retroviridae Proteins / Q000235:genetics
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral; D012191:Retroviridae Proteins; D015688:Oncogene Protein pp60(v-src)