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首页> 美国卫生研究院文献>Molecular and Cellular Biology >Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability.
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Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability.

机译：从酿酒酵母中鉴定和分离编码核糖核苷酸还原酶小亚基的基因：有丝分裂生存力所需的DNA损伤诱导基因。

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摘要

Ribonucleotide reductase catalyzes the first step in the pathway for the production of deoxyribonucleotides needed for DNA synthesis. The gene encoding the small subunit of ribonucleotide reductase was isolated from a Saccharomyces cerevisiae genomic DNA expression library in lambda gt11 by a fortuitous cross-reaction with anti-RecA antibodies. The cross-reaction was due to an identity between the last four amino acids of each protein. The gene has been named RNR2 and is centromere linked on chromosome X. The nucleotide sequence was determined, and the deduced amino acid sequence, 399 amino acids, shows extensive homology with other eucaryotic ribonucleotide reductases. Transplason mutagenesis was used to disrupt the RNR2 gene. A novel assay using colony color sectoring was developed to demonstrate visually that RNR2 is essential for mitotic viability. RNR2 encodes a 1.5-kilobase mRNA whose levels increase 18-fold after treatment with the DNA-damaging agent 4-nitroquinoline 1-oxide. CDC8 was also found to be inducible by DNA damage, but POL1 and URA3 were not inducible by 4-nitroquinoline 1-oxide. The expression of these genes defines a new mode of regulation for enzymes involved in DNA biosynthesis and sharpens our picture of the events leading to DNA repair in eucaryotic cells.
机译：核糖核苷酸还原酶催化DNA合成所需脱氧核糖核苷酸生产途径的第一步。通过与抗RecA抗体的偶然交叉反应从λgt11中的酿酒酵母基因组DNA表达文库中分离出编码核糖核苷酸还原酶小亚基的基因。交叉反应归因于每种蛋白质的最后四个氨基酸之间的同一性。该基因已命名为RNR2，在X染色体上与着丝粒相连。确定了核苷酸序列，推导的氨基酸序列399个氨基酸与其他真核生物核糖核苷酸还原酶具有广泛的同源性。使用Transplason诱变来破坏RNR2基因。开发了一种使用菌落颜色分区的新颖测定法，以在视觉上证明RNR2对于有丝分裂生存力至关重要。 RNR2编码一个1.5碱基碱基的mRNA，在用DNA破坏剂4-硝基喹啉1-氧化物处理后其水平增加18倍。还发现CDC8可被DNA损伤诱导，但POL1和URA3不能被4-硝基喹啉1氧化物诱导。这些基因的表达为参与DNA生物合成的酶定义了一种新的调节模式，并使我们对导致真核细胞DNA修复的事件的认识更加清晰。

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