<TEX>$ski^-$</TEX> 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자

A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in <TEX>$ski^-$</TEX> Host Cells

The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and <TEX>$M_{1}$</TEX>, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, <TEX>$ski^{-}$</TEX> mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in <TEX>$ski^{-}$</TEX> cells (not in <TEX>$SKI^{+}$</TEX> cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.