ANALYSIS OF SPONTANEOUS FRAMESHIFT MUTATIONS IN REV1 AND REV1-1 STRAINS OF SACCHAROMYCES-CEREVISIAE
ANALYSIS OF SPONTANEOUS FRAMESHIFT MUTATIONS IN REV1 AND REV1-1 STRAINS OF SACCHAROMYCES-CEREVISIAE
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DOI：
10.1016/0027-5107(95)00064-p
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发表时间：
1995-09-01
影响因子：
2.3
通讯作者：
PLEWA, MJ
中科院分区：
文献类型：
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作者：
KALINOWSKI, DP;LARIMER, FW;PLEWA, MJ
移码突变通过多种机制发生。为了更好地了解这些机制的本质，我们确定了酿酒酵母 REV1 和 rev1-1 菌株的 HIS4 基因中 232 个独立的自发移码突变的 DNA 序列变化。所有移码突变体都是根据其恢复 +1 移码突变 his4-38 的能力来选择的。 DNA 序列信息使用两种方法恢复：质粒 pMP4 的双链缺口修复和聚合酶链式反应 (PCR)。使用这些技术，生成了 his4-38 自发回复的饱和突变谱。两种菌株中最常发生的突变事件是-1移码，但也观察到+2移码、更大的缺失、更大的插入和更复杂的突变。在 REV1 和 rev1-1 菌株之间，我们注意到 -1 移码突变的分布存在显着差异。此外，虽然对于-1移码事件，通过双链间隙修复或PCR确定的回复谱之间没有显着差异，但通过双链间隙修复恢复的移码突变类型（仅-1移码和1个+2移码）与使用PCR恢复的移码突变类型（-1移码、+2移码、更大的删除和插入以及更复杂的突变）之间存在令人惊讶的显着差异。这种差异可能反映了双链断裂修复固有的选择机制，避免了包括复杂改变的染色体序列。
Frameshift mutations occur by a number of mechanisms. To better understand the nature of these mechanisms, we determined the DNA sequence changes of 232 independent, spontaneous frameshift mutations in the HIS4 gene of REV1 and rev1-1 strains of Saccharomyces cerevisiae. All frameshift mutants were selected based on their ability to revert the +1 frameshift mutation his4-38. DNA sequence information was recovered using two approaches - the double-strand gap repair of plasmid pMP4, and the polymerase chain reaction (PCR). Using these techniques, saturated mutation spectra for the spontaneous reversion of his4-38 were generated. The most frequently occurring mutational events in both strains were -1 frameshifts, but +2 frameshifts, larger deletions, larger insertions and more complex mutations were also observed. Between the REV1 and rev1-1 strains, we noticed a significant difference in the distribution of -1 frameshift mutations. In addition, while for -1 frameshift events there was no significant difference between the reversion spectra determined by double-strand gap repair or PCR, there was a surprisingly significant difference between the types of frameshift mutations recovered by double-strand gap repair (only -1 frameshifts and one +2 frameshift), and those recovered using PCR (-1 frameshifts, +2 frameshifts, larger deletions and insertions, and more complex mutations). This difference may reflect a selectional mechanism inherent in double-strand break repair that avoids chromosomal sequences which include complex alterations.