Lichten M, Goyon C, Schultes N P, Treco D, Szostak J W, Haber J E, Nicolas A
Laboratory of Biochemistry, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1990 Oct;87(19):7653-7. doi: 10.1073/pnas.87.19.7653.
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.
我们利用变性凝胶电泳对酵母减数分裂产物中的异源双链DNA进行了物理展示。我们检测了arg4 - nsp处的异源双链形成情况,arg4 - nsp是一个G.C到C.G的颠换,表现出中等程度的减数分裂后分离水平。在两种可能的arg4 - nsp/ARG4错配(G.G和C.C)中,在错配修复能力正常(Pms1 +)的二倍体产生的孢子中仅检测到C.C。相比之下,在Pms1 - 二倍体产生的孢子中,C.C和G.G的含量几乎相等。这些结果证实了先前的推测,即减数分裂后分离的孢子在相关标记位点含有异源双链DNA,C.C的修复频率低于G.G,并且PMS1基因产物在错配校正中发挥作用。结合Pms1 + ARG4/arg4 - nsp二倍体产生的3 +:5m(野生型:突变型)四分体(+,+,+/m,m)比5 +:3m四分体(+,+/m,m,m)多3倍的观察结果,这些结果表明,在减数分裂过程中,ARG4处异源双链DNA的形成涉及DNA双链有义(非转录)链的优先转移。