DNA Recombination of Thermophiles

嗜热菌 DNA 重组

• 批准号: 11694207
• 负责人: KURAMITSU Seiki
• 金额: $ 5.57万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694207/
• 关键词: homologous recombination; DNA repair; extreme thermophile; hyperthermophile; archae; RecA; RadA; DNA; 遺伝子組換え; Thermus thermophilus HB8; Desulfurcoccus amyloliticus

In living organisms, genetic recombination is related to the DNA damage. DNA damage often arises as a result of genetic recombination, DNA replication, and other processes. In prokaryotes, about fifty enzymes participate in the DNA repair reactions. From thermophilic bacteria including hyperthermophile, we have cloned the DNA-repair related genes, which include recombinational repair enzymes (RecA, RuvA, RuvB, RuvC, ssb), base excision repair enzymes (MutM, MutY, EndoIII, EndoV), nucleotide excision repair enzymes (UvrA, UvrB, UvrC, UvrD), mismatch repair-enzymes (MutS, MutL), photolyase, and DNA polymerase I and ligase. Most of these DNA repair systems are common to many organisms, including those of Homo sapiens. These thermophilic enzymes, as expected, were stable. It was also found that the domains of the respective proteins can be easily isolated by partial digestion with protease. Some repair enzymes, including MutM, UvrB, and photolyase, have been crystallized and their three-dimensional structures have been determined. These thermophilic enzymes have been adapted to high temperatures. Some hyperthermophilic repair enzymes dramatically changed its conformation and increased the activity above 75℃

在生物体中，基因重组与DNA损伤有关。 DNA 损伤通常是由基因重组、DNA 复制和其他过程引起的。在原核生物中，大约有 50 种酶参与 DNA 修复反应。我们从嗜热菌（包括超嗜热菌）中克隆了DNA修复相关基因，包括重组修复酶（RecA、RuvA、RuvB、RuvC、ssb）、碱基切除修复酶（MutM、MutY、EndoIII、EndoV）、核苷酸切除修复酶（UvrA、UvrB、UvrC、 UvrD)、错配修复酶 (MutS、MutL)、光裂合酶、DNA 聚合酶 I 和连接酶。这些 DNA 修复系统大多数是许多生物体所共有的，包括智人。正如预期的那样，这些嗜热酶是稳定的。还发现通过用蛋白酶部分消化可以容易地分离各个蛋白质的结构域。一些修复酶，包括MutM、UvrB和光裂合酶，已经结晶并确定了它们的三维结构。这些嗜热酶已适应高温。一些超高温修复酶在75℃以上构象发生急剧改变，活性增强

项目成果

Sugahara, M.: "Crystal Structure of a Repair Enzyme of Oxidatively-Damaged-DNA, MutM (Fpg), from an Extreme Thermophile, Thermus thermophilus HB8"EMBO J.. 19 (15). 3857-3869 (2000)

Sugahara, M.：“来自极端嗜热菌、嗜热栖热菌 HB8 的氧化损伤 DNA 修复酶 MutM (Fpg) 的晶体结构”EMBO J.. 19 (15)。

Sugahara,M.: "Crystal Structure of a Repair Enzyme of Oxidatively-Damaged-DNA, MutM (Fpg), from an Extreme Thermophile, Thermus thermophilus HB8"EMBO J.. 19(15). 3857-3869 (2000)

Sugahara,M.：“氧化损伤 DNA 修复酶 MutM (Fpg) 的晶体结构，来自极端嗜热菌，嗜热栖​​热菌 HB8”EMBO J.. 19(15)。

Isaev-Ivanov,V.V.: "Fluorescence and Excitation Escherichia coli RecA Protein Spectra Analyzed Separately for Tyrosine and Tryptophan Residues"Arch.Biochem.Biophys.. 376(1). 124-140 (2000)

Isaev-Ivanov，V.V.：“分别分析酪氨酸和色氨酸残基的荧光和激发大肠杆菌 RecA 蛋白光谱”Arch.Biochem.Biophys.. 376(1)。

Nakagawa,N.: "Crystal Structure of Thermus thermophilus HB8 UvrB Protein,A Key Enzyme of Nucleotide Excision Repair"J.Biochem.. 126(6). 986-990 (1999)

Nakakawa,N.：“嗜热栖热菌 HB8 UvrB 蛋白的晶体结构，核苷酸切除修复的关键酶”J.Biochem.. 126(6)。

Sugahara,M.: "Crystal Structure of a Repair Enzyme of Oxidatively-Damaged-DNA,MutM (Fpg) , from an Extreme Thermophile, Thermus thermophilus HB8"EMBO J.. 19(15). 3857-3869 (2000)

Sugahara,M.：“氧化损伤 DNA 修复酶 MutM (Fpg) 的晶体结构，来自极端嗜热菌，嗜热栖​​热菌 HB8”EMBO J.. 19(15)。