Mutational Analysis of Conserved Hydrophobic Amino Acid Residues in the N-Terminal Region of DnaA Protein
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概要
- 論文の詳細を見る
DnaA is the initiator of chromosomal DNA replication in F. coli. We previously reported that conserved hydrophobic amino acid residues in the N-terminal region of DnaA (126 and L40) are essential for DNA replication in vivo and in vitro using mutant DnaA proteins (DnaAI26S and DnaAL40S). In this study, we introduced further random mutations to find intragenic suppressors for dnaAI26S or dnaAL40S. By direct DNA sequence, a mutation which causes substitution of the Ser (Ile, in the wild-type DnaA) with Phe (DnaAI26F or DnaAL40F) was found in all of the suppressors. Site-directed mutational analysis showed that DnaAI26L, and DnaAL40I, but not DnaAI26S or DnaAL40S, were active for oriC DNA replication in cells. Furthermore, purified DnaAI26F but not DnaAI26S was active for oriC DNA replication in a crude extract. These results strongly suggest that hydrophobic amino acid residues in these positions of DnaA (126 and L40) are important for the function of this protein as an initiator of DNA replication both in vivo and in vitro.
- 公益社団法人日本薬学会の論文
- 2002-07-01
著者
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土屋 照雄
Gifu Prefectural Hospital
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Tsuchiya Tomofusa
岡山大学 薬学部
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Tsuchiya Tomofusa
Faculty Of Pharmaceutical Sciences Okayama University
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Mizushima Tohru
Graduate School of Medical and Pharmaceutical Sciences, Kumamoto University
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Mima Takehiko
岡山大学 医歯薬学総合研究科
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Miki T
Faculty Of Pharmaceutical Sciences Kyushu University
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Tsuchiya T
Faculty Of Pharmaceutical Sciences Okayama University
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Tsuchiya T
Okayama Univ. Okayama
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MIZUSHIMA Tohru
Faculty of Pharmaceutical Sciences, Okayama University
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Koterasawa Motohiro
Department Of Molecular Microbiology Graduate School Of Medicine Dentistry And Pharmaceutical Scienc
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Koterasawa Motohiro
Faculty Of Pharmaceutical Sciences Okayama University
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Makise Masaki
Faculty of Pharmaceutical Sciences, Okayama University
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Makise Masaki
Graduate School Of Medical And Pharmaceutical Sciences Kumamoto University
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Tsuchiya T
Faculty Of Pharmaceutical Sciences Hokuriku University
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Mizushima Tohru
Department Of Molecular Microbiology Graduate School Of Medicine Dentistry And Pharmaceutical Scienc
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Mizushima Tohru
Faculty Of Pharmaceutial Sciences Kyushu University
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Mizushima Tohru
Faculty Of Engineering. Tokai University
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MIMA Shinji
Faculty of Pharmaceutical Sciences, Okayama University
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Tsuchiya Tomofusa
Department Of Microbiology Fujita Health University
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Mizushima T
Faculty Of Pharmaceutial Sciences Kyushu University
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Makise Masaki
Faculti of Pharmaceutical Sciences Okairama University
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Mizushima Tohru
Faculti of Pharmaceutical Sciences Okairama University
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