The mcm2 mutation of yeast affects replication, rather than segregation or amplification of the two micron plasmid.
We have studied the maintenance of the endogenous two micron (2 mu) plasmid in a strain of yeast carrying the nuclear mutation mcm2. This mutation, earlier shown to affect the maintenance of yeast minichromosomes in an ARS-dependent manner, also affected the copy number of the 2 mu plasmid. The effect was more pronounced at 35 degrees C leading to the elimination of the plasmid from the cells cultured at this temperature. The mutant cells could be efficiently cured of the circle by transformation with 2 mu ORI-carrying hybrid vectors, an observation consistent with the low copy number of the endogenous plasmid. A chromosomal revertant of this mutant for another ARS(ARS1) was found also to confer stability on the 2 mu ORI-carrying minichromosomes and had elevated levels of the endogenous plasmid. The mutation neither affected the segregation nor the amplification process mediated by site-specific recombination at FRT sites requiring the FLP gene-encoded protein action. ARS131C, an ARS that was unaffected in the mutant at 25 degrees C, could elevate the copy number of a 2 mu hybrid vector in the mutant cells. In view of these results, some aspects of segregation and copy number control of the endogeneous plasmid have been discussed. We propose that the mutation impairs the 2 mu ORI function, leading to its loss.
Maiti
A K
AK
Department of Biochemistry, Bose Institute, Calcutta, India.
Sinha
P
P
D002455:Cell Division; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis*; D005784:Gene Amplification; D009154:Mutation; D010957:Plasmids; D011336:Probability; D011995:Recombination, Genetic; D012093:Replicon; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D013696:Temperature
D004271:DNA, Fungal