Effects of genome position and the DNA damage checkpoint on the structure and frequency of sod2 gene amplification in fission yeast.
The Schizosaccharomyces pombe sod2 gene, located near the telomere on the long arm of chromosome I, encodes a Na+ (or Li+)/H+ antiporter. Amplification of sod2 has previously been shown to confer resistance to LiCl. We analyzed 20 independent LiCl-resistant strains and found that the only observed mechanism of resistance is amplification of sod2. The amplicons are linear, extrachromosomal elements either 225 or 180 kb long, containing both sod2 and telomere sequences. To determine whether proximity to a telomere is necessary for sod2 amplification, a strain was constructed in which the gene was moved to the middle of the same chromosomal arm. Selection of LiCl-resistant strains in this genetic background also yielded amplifications of sod2, but in this case the amplified DNA was exclusively chromosomal. Thus, proximity to a telomere is not a prerequisite for gene amplification in S. pombe but does affect the mechanism. Relative to wild-type cells, mutants with defects in the DNA damage aspect of the rad checkpoint control pathway had an increased frequency of sod2 amplification, whereas mutants defective in the S-phase completion checkpoint did not. Two models for generating the amplified DNA are presented.
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030, USA.
T E
TE
Patterson
E B
EB
Albrecht
P
P
Nurse
S
S
Sazer
G R
GR
Stark
D002453:Cell Cycle / Q000235:genetics; D002874:Chromosome Mapping; D004249:DNA Damage / Q000235:genetics*; D005784:Gene Amplification; D005787:Gene Frequency; D005800:Genes, Fungal; D016681:Genome, Fungal; D009154:Mutation; D012568:Schizosaccharomyces / Q000235:genetics*; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics*; D016615:Telomere / Q000235:genetics
D017923:Sodium-Hydrogen Exchangers