Effects of excess centromeres and excess telomeres on chromosome loss rates.
The linear chromosomes of eukaryotes contain specialized structures to ensure their faithful replication and segregation to daughter cells. Two of these structures, centromeres and telomeres, are limited, respectively, to one and two copies per chromosome. It is possible that the proteins that interact with centromere and telomere DNA sequences are present in limiting amounts and could be competed away from the chromosomal copies of these elements by additional copies introduced on plasmids. We have introduced excess centromeres and telomeres into Saccharomyces cerevisiae and quantitated their effects on the rates of loss of chromosome III and chromosome VII by fluctuation analysis. We show that (i) 600 new telomeres have no effect on chromosome loss; (ii) an average of 25 extra centromere DNA sequences increase the rate of chromosome III loss from 0.4 x 10(-4) events per cell division to 1.3 x 10(-3) events per cell division; (iii) centromere DNA (CEN) sequences on circular vectors destabilize chromosomes more effectively than do CEN sequences on 15-kb linear vectors, and transcribed CEN sequences have no effect on chromosome stability. We discuss the different effects of extra centromere and telomere DNA sequences on chromosome stability in terms of how the cell recognizes these two chromosomal structures.
Runge
K W
KW
Fred Hutchinson Cancer Research Center, Division of Basic Sciences, Seattle, Washington 98104.
Wellinger
R J
RJ
Zakian
V A
VA
D002455:Cell Division; D002503:Centromere / Q000502:physiology*; D015825:Chromosomes, Fungal / Q000502:physiology*; D003001:Cloning, Molecular / Q000379:methods; D004274:DNA, Recombinant / Q000378:metabolism; D004171:Diploidy; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development
D004274:DNA, Recombinant