Nag D K, Kurst A
Department of Biomedical Sciences, School of Public Health, State University of New York, Albany 12201, USA.
Genetics. 1997 Jul;146(3):835-47. doi: 10.1093/genetics/146.3.835.
Palindromic sequences have the potential to form hairpin or cruciform structures, which are putative substrates for several nucleases and mismatch repair enzymes. A genetic method was developed to detect such structures in vivo in the yeast Saccharomyces cerevisiae. Using this method we previously showed that short hairpin structures are poorly repaired by the mismatch repair system in S. cerevisiae. We show here that mismatches, when present in the stem of the hairpin structure, are not processed by the repair machinery, suggesting that they are treated differently than those in the interstrand base-paired duplex DNA. A 140-bp-long palindromic sequence, on the contrary, acts as a meiotic recombination hotspot by generating a site for a double-strand break, an initiator of meiotic recombination. We suggest that long palindromic sequences undergo cruciform extrusion more readily than short ones. This cruciform structure then acts as a substrate for structure-specific nucleases resulting in the formation of a double-strand break during meiosis in yeast. In addition, we show that residual repair of the short hairpin structure occurs in an MSH2-independent pathway.
回文序列有可能形成发夹或十字形结构,这些结构是几种核酸酶和错配修复酶的假定底物。我们开发了一种遗传方法来在酿酒酵母体内检测此类结构。利用这种方法,我们之前表明,短发夹结构在酿酒酵母中很难被错配修复系统修复。我们在此表明,当错配存在于发夹结构的茎中时,修复机制不会对其进行处理,这表明它们与链间碱基配对的双链DNA中的错配处理方式不同。相反,一个140个碱基对长的回文序列通过产生一个双链断裂位点(减数分裂重组的起始点)而作为减数分裂重组热点。我们认为,长回文序列比短回文序列更容易发生十字形突出。这种十字形结构随后作为结构特异性核酸酶的底物,导致酵母减数分裂期间形成双链断裂。此外,我们表明,短发夹结构的残余修复发生在一条不依赖MSH2的途径中。