| Abstract: | A mutation in polyomavirus large T antigen which affects viral DNA synthesis was discovered in strain NG59RA (RA). The effect was most visible in nonpermissive cells. Although a substantial yield in DNA synthesis is normally observed in infections of Fischer rat cells when these are maintained at 33 degrees C (D.L. Hacker, K.H. Friderici, C. Priehs, S. Kalvonjian, and M.M. Fluck, p. 173-181, in R.E. Moses and W.C. Summers, ed., DNA Replication and Mutagenesis, 1988; D.L. Hacker and M.M. Fluck, Mol. Cell. Biol., in press), a 10- to 20-fold decrease in yield was obtained in infections with RA. The yield of free viral DNA in RA transformants was also strongly diminished, whether the transformants were maintained at 37 or 33 degrees C. A large reduction in the apparent number of integration sites, as well as a small reduction in the incidence of tandem integration of the viral genome, was observed in F-111 or FR-3T3 cells transformed by the mutant strain. This appears not to be directly related to the number of integration templates. A DNA fragment was identified which rescues these phenotypes. The fragment is located between the HindIII and NsiI restriction sites (nucleotides 1656 to 1910), a region which encodes only large T antigen. Sequence analysis of this region reveals a C-to-G transition at nucleotide 1791 which causes a proline-to-alanine change in the amino acid sequence of large T antigen. No other mutations have been previously reported in this region of large T antigen. |
