Leem S H, Ropp P A, Sugino A
Department of Molecular Immunology, Osaka University, Japan.
Nucleic Acids Res. 1994 Aug 11;22(15):3011-7. doi: 10.1093/nar/22.15.3011.
We identified and purified a new DNA polymerase (DNA polymerase IV), which is similar to mammalian DNA polymerase beta, from Saccharomyces cerevisiae and suggested that it is encoded by YCR14C (POLX) on chromosome III. Here, we provided a direct evidence that the purified DNA polymerase IV is indeed encoded by POLX. Strains harboring a pol4 deletion mutation exhibit neither mitotic growth defect nor a meiosis defect, suggesting that DNA polymerase IV participates in nonessential functions in DNA metabolism. The deletion strains did not exhibit UV-sensitivity. However, they did show weak sensitivity to MMS-treatment and exhibited a hyper-recombination phenotype when intragenic recombination was measured during meiosis. Furthermore, MAT alpha pol4 delta segregants had a higher frequency of illegitimate mating with a MAT alpha tester strain than that of wild-type cells. These results suggest that DNA polymerase IV participates in a double-strand break repair pathway. A 3.2kb of the POL4 transcript was weakly expressed in mitotically growing cells. During meiosis, a 2.2 kb POL4 transcript was greatly induced, while the 3.2 kb transcript stayed at constant levels. This induction was delayed in a swi4 delta strain during meiosis, while no effect was observed in a swi6 delta strain.
我们从酿酒酵母中鉴定并纯化出一种新的DNA聚合酶(DNA聚合酶IV),它与哺乳动物的DNA聚合酶β相似,并推测它由III号染色体上的YCR14C(POLX)编码。在此,我们提供了直接证据,证明纯化的DNA聚合酶IV确实由POLX编码。携带pol4缺失突变的菌株既没有有丝分裂生长缺陷,也没有减数分裂缺陷,这表明DNA聚合酶IV参与DNA代谢中的非必需功能。缺失菌株对紫外线不敏感。然而,它们对MMS处理表现出较弱的敏感性,并且在减数分裂期间测量基因内重组时表现出超重组表型。此外,MATα pol4δ分离株与MATα测试菌株发生非法交配的频率高于野生型细胞。这些结果表明DNA聚合酶IV参与双链断裂修复途径。3.2kb的POL4转录本在有丝分裂生长的细胞中弱表达。在减数分裂期间,2.2kb的POL4转录本被大量诱导,而3.2kb的转录本保持恒定水平。在减数分裂期间,这种诱导在swi4δ菌株中延迟,而在swi6δ菌株中未观察到影响。