Detection of heteroduplex DNA molecules among the products of Saccharomyces cerevisiae meiosis.
We have used denaturant-gel electrophoresis to provide a physical demonstration of heteroduplex DNA in the products of yeast meiosis. We examined heteroduplex formation at arg4-nsp, a G.C----C.G transversion that displays a moderately high level of postmeiotic segregation. Of the two possible arg4-nsp/ARG4 mismatches (G.G and C.C), only C.C was detected in spores from mismatch repair-competent (Pms1+) diploids. In contrast, C.C and G.G were present at nearly equal levels in spores from Pms1- diploids. These results confirm previous suggestions that postmeiotic segregation spores contain heteroduplex DNA at the site of the marker in question, that C.C is repaired less frequently than is G.G, and that the PMS1 gene product plays a role in mismatch correction. Combined with the observation that Pms1+ ARG4/arg4-nsp diploids produce 3 times more 3+:5m (wildtype:mutant) tetrads (+, +, +/m, m) than 5+:3m tetrads (+, +/m, m, m), these results indicate that, during meiosis, formation of heteroduplex DNA at ARG4 involves preferential transfer of the sense (nontranscribed) strand of the DNA duplex.
Lichten
M
M
Laboratory of Biochemistry, National Institutes of Health, Bethesda, MD 20892.
Goyon
C
C
Schultes
N P
NP
Treco
D
D
Szostak
J W
JW
Haber
J E
JE
Nicolas
A
A
D001483:Base Sequence; D003433:Crosses, Genetic; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics*; D005838:Genotype; D008540:Meiosis; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D013172:Spores, Fungal / Q000502:physiology
D004271:DNA, Fungal; D009692:Nucleic Acid Heteroduplexes; D015345:Oligonucleotide Probes