DETECTION OF HETERODUPLEX DNA-MOLECULES AMONG THE PRODUCTS OF SACCHAROMYCES-CEREVISIAE MEIOSIS

被引:98
作者
LICHTEN, M
GOYON, C
SCHULTES, NP
TRECO, D
SZOSTAK, JW
HABER, JE
NICOLAS, A
机构
[1] BRANDEIS UNIV,ROSENSTIEL BASIC MED SCI RES CTR,WALTHAM,MA 02254
[2] UNIV PARIS 11,INST GENET & MICROBIOL,F-91405 ORSAY,FRANCE
[3] MASSACHUSETTS GEN HOSP,DEPT MOLEC BIOL,BOSTON,MA 02114
关键词
denaturant-gel electrophoresis; mismatch repair; postmeiotic segregation; preferential strand transfer; yeast;
D O I
10.1073/pnas.87.19.7653
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:7653 / 7657
页数:5
相关论文
共 29 条
[1]   THE NUCLEOTIDE-SEQUENCE OF THE YEAST ARG4 GENE [J].
BEACHAM, IR ;
SCHWEITZER, BW ;
WARRICK, HM ;
CARBON, J .
GENE, 1984, 29 (03) :271-279
[2]   SPECIFICITY OF MISMATCH REPAIR FOLLOWING TRANSFORMATION OF SACCHAROMYCES-CEREVISIAE WITH HETERODUPLEX PLASMID DNA [J].
BISHOP, DK ;
ANDERSEN, J ;
KOLODNER, RD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (10) :3713-3717
[3]   THE ROLE OF HETERODUPLEX CORRECTION IN GENE CONVERSION IN SACCHAROMYCES-CEREVISIAE [J].
BISHOP, DK ;
WILLIAMSON, MS ;
FOGEL, S ;
KOLODNER, RD .
NATURE, 1987, 328 (6128) :362-364
[4]   A PATHWAY FOR GENERATION AND PROCESSING OF DOUBLE-STRAND BREAKS DURING MEIOTIC RECOMBINATION IN SACCHAROMYCES-CEREVISIAE [J].
CAO, L ;
ALANI, E ;
KLECKNER, N .
CELL, 1990, 61 (06) :1089-1101
[5]   GENE CONVERSION, RECOMBINATION NODULES, AND THE INITIATION OF MEIOTIC SYNAPSIS [J].
CARPENTER, ATC .
BIOESSAYS, 1987, 6 (05) :232-236
[6]   A TECHNIQUE FOR RADIOLABELING DNA RESTRICTION ENDONUCLEASE FRAGMENTS TO HIGH SPECIFIC ACTIVITY [J].
FEINBERG, AP ;
VOGELSTEIN, B .
ANALYTICAL BIOCHEMISTRY, 1983, 132 (01) :6-13
[7]   DNA FRAGMENTS DIFFERING BY SINGLE BASE-PAIR SUBSTITUTIONS ARE SEPARATED IN DENATURING GRADIENT GELS - CORRESPONDENCE WITH MELTING THEORY [J].
FISCHER, SG ;
LERMAN, LS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (06) :1579-1583
[8]  
FOGEL S, 1981, MOL BIOL YEAST SACCH, P289
[9]   SOME FEATURES OF GENETIC-RECOMBINATION IN PROCARYOTES [J].
FOX, MS .
ANNUAL REVIEW OF GENETICS, 1978, 12 :47-68
[10]  
HASTINGS P J, 1987, P139