| Project/Area Number |
05044132
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Osaka University |
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Principal Investigator |
OGAWA Hideyuki Faculty of Science, Osaka University, 理学部, 教授 (70028207)
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| Co-Investigator(Kenkyū-buntansha) |
YURI V.Kil ペテルスブルグ 核物理学研究所, 分子放射線生物学部門・分子遺伝学研究室, 研究員
ALEXSEEV Andrej a. St.Petersburg Nuclear Physics Institute in Russia, 分子遺伝学部門, 研究員
LANZOV Vladi ペテルスブルグ 核物理学研究所, 分子遺伝学部門, 部長
KURAMITSU Seiki Faculty of Science, 理学部, 教授 (60153368)
ITOH Tateo Faculty of Science, 理学部, 助教授 (40051817)
OGAWA Tomoko Faculty of Science, 理学部, 講師 (80028208)
LANZOV Vladislav a. St.Petersburg Nuclear Physics Institute in Russia
KIL Yuri v. St.Petersburg Nuclear Physics Institute in Russia
MINICASIMOVI イバチュリン ファ ペテルスブルグ核物理学研究所, 分子放射線生物学部門, 研究員
KONSTANTINOV カボエフ オレ ペテルスブルグ核物理学研究所, 分子放射線生物学部門, 室長
NIKITCHNA Zl ペテルスブルグ物性物理学研究所, 室長
ALEXSEEVICH アレキセエフ アント ペテルスブルグ核物理学研究所, 分子遺伝学, 研究員
ALEXSANDROVI ランゾフ ウ ペテルスブルグ核物理学研究所, 分子遺伝学, 部長
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | the recA gene of thermophilic bacteria / RecA2278-5 / St.Petersburg Nuclear Physics Institute in Russia / temperature sensitive RecA protein / 温度感受性recA2278-5変異 / RecAのα-ヘリックスHの変異 / 好熱性菌のコドン使用頻度 / ATP結合蛋白質の共通アミノ酸配列 / RecA蛋白質 / 好熱菌recA遺伝子のクローニング / 熱安定性RecA蛋白質 / 組換え機構の普遍性 / RecA蛋白質の構造と機能の関連 |
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| Research Abstract |
Among important problems in recombination, one of the hottest problems is to reveal a relationship between structure and function of RecA protein.To pursue this problem, it is inevitable to have suitable mutants in the recA gene.We learned that having many interesting recA mutants in phenotype, Dr.Lanzov, St.Petersburg Nuclear Physics Institute in Russia, was not familiar in physico-chemical analysis of RecA protein.Therefore we agreed to do a joint work using his useful recA mutants.Furthermore, he also has a big collection of various kinds of thermophilic bacteria, proteins of which are stable enough for sever conditions during physico-chemical analysis.We also agreed to isolate a suitable recA gene from a thermophilic bacteria to provide a heat stable RecA protein to facilitate physico-chemical analyzes.Dr.Lanzov with his colleagues came to our lab and did the joint works for these two years and obtained following results.
RecA2278-5 is a mutant RecA protein bearing two amino acid substitutions, Gly278 by Thr and Val275 by Phe, in the a-helix H of C-terminal sub-domain of the RecA.The mutant is defective in genetic recombination and SOS-repair at 42*C.We compared the biochemical activities of RecA2278-5 with those of a wild type RecA protein at 32ﾟand 42ﾟC.The thermosensitive multiple deficiencies of the RecA2278-5 protein suggest that the structural stability of C-subdomain of the RecA protein is necessary for RecA-ATP-ssDNA helix filament formation that is a primary step of homologous recombination in bacteria.
The recA gene of a thermophilic bacteria was successfully isolated by PCR methods, in which the primers were designed after a common amino acid sequence for an ATP binding domain of RecA proteins.Nucleotide sequence analysis revealed that most of the third nucleotide in a codon is G or C.Therefore the DNA is GC rich and heat-stable. This characteristic is quite reasonable for thermophilic bacteria.
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