Mutation near the polyoma DNA replication origin permits productive infection of F9 embryonal carcinoma cells.
F9 mouse embryonal carcinoma cells are resistant to productive infection by wild-type polyoma virus. Continued passage of F9 cells initially infected with wild-type polyoma virus eventually leads to the selection of polyoma virus mutants that are capable of productive infection of undifferentiated F9 cells. Three mutants, PyF101, PyF111 and PyF441, have been plaque-purified and examined. All three PyF mutant DNAs are altered from the wild-type sequence in the Pvu II-4 fragment that spans 67.6 to 70.2 map units on the polyoma genome. PyF441 has a single base change of A to G at 69.6 map units. PyF101 and PyF111 DNAs also contain this point mutation at 69.6 map units. In addition, PyF101 and PyF111 DNAs have exact tandem duplications of 54 and 31 bp, respectively, of sequences encompassing the point mutation, and both copies of the tandem duplication have the point mutation. Other than these changes, no difference exists in the nucleotide sequences of wild-type and PyF mutant DNAs from the BcI I site at 65.6 map units clockwise through the origin of viral DNA replication to the BgI I site at 72.2 map units. DNA infections of F9 cells with wild-type-mutant hybrid DNAs formed by ligation of heterologous combinations of the small and large DNA fragments generated by double digestion with the restriction enzymes BcII and BGI I show that the DNA sequence changes described above are responsible for the ability of the PyF mutants to infect F9 cells.
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics; D000956:Antigens, Viral / Q000235:genetics; D000957:Antigens, Viral, Tumor; D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation; D051379:Mice; D009154:Mutation; D011120:Polyomavirus / Q000235:genetics*; D013724:Teratoma / Q000382:microbiology*; D014779:Virus Replication
D000951:Antigens, Neoplasm; D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D004279:DNA, Viral