FUKUI Kenji
| Faculty Division of Human Life and Environmental Sciences Research Group of Food Science and Nutrition | Associate Professor |
Last Updated :2026/02/19
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Name (Japanese)
FukuiName (Kana)
Kenji
Research Interests
Research Areas
Research History
- Apr. 2025 - Present, Osaka Medical and Pharmaceutical University, 医学部 生化学教室, 非常勤講師 (兼任)
- Apr. 2025 - Present, Nara Women's University, Faculty of Human Life and Environment Department of Food Science and Nutrition, Associate Professor
- Oct. 2024 - Mar. 2025, Nara Women's University, 研究院 生活環境科学系 食物栄養学領域, 非常勤講師 (兼任)
- Apr. 2019 - Mar. 2025, Baika Women's University, 看護保健学部 看護学科, 非常勤講師 (兼任)
- Apr. 2014 - Mar. 2025, Osaka Medical and Pharmaceutical University, Faculty of Medicine, Department of Biochemistry, Assistant Professor
- Apr. 2023 - Mar. 2024, Osaka Metropolitan University, 理学部, 非常勤講師 (兼任)
- Apr. 2013 - Mar. 2016, Osaka University, 理学研究科, 招へい研究員 (兼任)
- Apr. 2013 - Mar. 2014, Transgenic Inc., Researcher
- Nov. 2012 - Mar. 2013, Osaka University, 産学連携本部, Specially Appointed Assistant Professor
- Apr. 2007 - Oct. 2012, RIKEN, SPring-8 Center, Researcher
- Apr. 2005 - Mar. 2007, JSPS, Fellow DC2
Education
Professional Memberships
■Ⅱ．研究活動実績
Published Papers
- Refereed, Nature Communications, KH–R3H domain cooperation in RNA recognition by the global RNA-binding protein KhpB, Kenji Fukui; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Takato Yano, 03 Sep. 2025, 16, 8028, Scientific journal, 10.1038/s41467-025-62302-y
- Refereed, The Journal of Biochemistry, The crystal structure of the small zinc-finger protein ZifS from Thermus thermophilus HB8., Saki Kurinami; Kenji Fukui; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Hiroki Okanishi; Yoshikatsu Kanai; Takato Yano; Ryoji Masui, Zinc finger domains are important interaction modules for binding to nucleic acids, proteins, lipids and small molecules. Many small-sized zinc finger proteins are encoded in bacterial genomes, but most of them have not been functionally annotated. We focused on TTHA0897, ZifS, as a small zinc finger protein from the extremely thermophilic eubacterium Thermus thermophilus HB8. In vivo experiments suggested that the cellular function of ZifS is related to the growth transition of T. thermophilus from the lag to the exponential phase under nutritionally limited conditions. In vitro biochemical experiments, including electrophoretic mobility shift assay and pull-down assay, yielded no clues about molecular functions of ZifS. X-ray crystallographic analysis revealed that the dimeric ZifS globally forms a cylinder-like structure, although ZifS dimer has no overall structural similarity to other known zinc finger proteins. The zinc ion-binding manner of ZifS fitted the characteristics of the zinc ribbon fold, which are mostly found in domains from proteins involved in the transcriptional and translational machinery. The crystal structure of ZifS is the first experimental insight into the molecular structure of this protein family, revealing several conserved features that may be functionally relevant., 31 Jul. 2025, 178, 2, 121, 133, Scientific journal, True, 10.1093/jb/mvaf028
- Refereed, Structure, ATP binding controls the molecular function of bacterial MutS2 by mediating closure of the dimeric clamp structure., Kenji Fukui; Takeshi Murakawa; Nobumasa Hino; Naoyuki Kondo; Takato Yano, MutS2 recognizes branched DNA structures to regulate homologous recombination. MutS2 also has a role in ribosome recycling, where it resolves collided ribosomes. These functions require ATP-dependent conformational changes of MutS2. In the known nucleotide-free and ADP-bound MutS2 structures the dimeric clamp-like structure adopts open conformations. Here, we present the crystal structure of MutS2 with a bound ATP analog revealing a closed conformation of the clamp. Experiments with MutS2, where an unnatural photo-crosslinking capable amino acid was introduced into the clamp revealed that ATP-dependent closure also occurs in solution. Binding of MutS2 to a terminal-containing DNA was not affected by ATP, whereas that to a terminal-less DNA was reinforced. These findings suggest that clamp closure enables MutS2 to stay bound to recombination intermediates, which might regulate recombination. Furthermore, closure of the clamp provides insights into the mechanism of dissociation of collided ribosomes mediated by MutS2., 20 Mar. 2025, 33, 1007, 1015, Scientific journal, True, 10.1016/j.str.2025.03.003
- Refereed, Biomedicines, Nerve Enlargement in Patients with INF2 Variants Causing Peripheral Neuropathy and Focal Segmental Glomerulosclerosis., Quynh Tran Thuy Huong; Linh Tran Nguyen Truc; Hiroko Ueda; Kenji Fukui; Koichiro Higasa; Yoshinori Sato; Shinichi Takeda; Motoshi Hattori; Hiroyasu Tsukaguchi, Background: Charcot-Marie-Tooth (CMT) disease is an inherited peripheral neuropathy primarily involving motor and sensory neurons. Mutations in INF2, an actin assembly factor, cause two diseases: peripheral neuropathy CMT-DIE (MIM614455) and/or focal segmental glomerulosclerosis (FSGS). These two phenotypes arise from the progressive degeneration affecting podocytes and Schwann cells. In general, nerve enlargement has been reported in 25% of the demyelinating CMT subtype (CMT1), while little is known about the CMT-DIE caused by INF2 variants. Methods: To characterize the peripheral nerve phenotype of INF2-related CMT, we studied the clinical course, imaging, histology, and germline genetic variants in two unrelated CMT-DIE patients. Results: Patient 1 (INF2 p.Gly73Asp) and patient 2 (p.Val108Asp) first noticed walking difficulties at 10 to 12 years old. Both of them were electrophysiologically diagnosed with demyelinating neuropathy. In patient 2, the sural nerve biopsy revealed an onion bulb formation. Both patients developed nephrotic syndrome almost simultaneously with CMT and progressed into renal failure at the age of 16 to 17 years. Around the age of 30 years, both patients manifested multiple hypertrophy of the trunk, plexus, and root in the cervical, brachial, lumbosacral nerves, and cauda equina. The histology of the cervical mass in patient 2 revealed Schwannoma. Exome analysis showed that patient 2 harbors a germline LZTR1 p.Arg68Gly variant, while patient 1 has no schwannomatosis-related mutations. Conclusions: Peripheral neuropathy caused by INF2 variants may lead to the development of multifocal hypertrophy with age, likely due to the initial demyelination and subsequent Schwann cell proliferation. Schwannoma could co-occur when the tissues attain additional hits in schwannomatosis-related genes (e.g., LZTR1)., 08 Jan. 2025, 13, 1, Scientific journal, True, 10.3390/biomedicines13010127
- Refereed, PLoS One, The crystal structure of Thermus thermophilus UMP kinase complexed with a phosphoryl group acceptor and donor., Kenji Fukui; Anzu Nishiwaki; Noriko Nakagawa; Seiki Kuramitsu; Ryoji Masui, Nucleoside monophosphate kinases play crucial roles in biosynthesis and regeneration of nucleotides. Prokaryotic UMP kinase belongs to a family of amino acid kinases but not to other nucleoside monophosphate kinases. Although many structures of prokaryotic UMP kinase have been determined, limited structural information has been available on the conformational changes along the reaction and allosteric pathways. We determined the crystal structure of UMP kinase of an extreme thermophile Thermus thermophilus HB8 in ADP-UDP-bound form at 2.6-Å resolution. The structure of the ADP-UDP complex is the first structure of bacterial UMP kinase with a phosphoryl group donor and an acceptor. Upon simultaneous binding of ADP and UDP, the loop near ADP moved toward the active site without global open-closed conformational changes, compared to the ligand-free and UDP-bound forms. Such a shift was not observed for archaeal UMP kinases but had some similarities to those in other amino acid kinase families of enzymes., 2025, 20, 9, e0330398, Scientific journal, True, 10.1371/journal.pone.0330398
- Refereed, DNA Repair, Elsevier BV, The HNH endonuclease domain of the giant virus MutS7 specifically binds to branched DNA structures with single-stranded regions, Satoshi Yoshioka; Hirochika Kurazono; Koki Ohshita; Kenji Fukui; Masaharu Takemura; Shin-Ichiro Kato; Kouhei Ohnishi; Takato Yano; Taisuke Wakamatsu, Jan. 2025, 145, 103804, 103804, Scientific journal, 10.1016/j.dnarep.2024.103804
- Refereed, Protein Science, dUTP pyrophosphatases from hyperthermophilic eubacterium and archaeon: Structural and functional examinations on the suitability for PCR application., Kenji Fukui; Naoyuki Kondo; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Takato Yano, Deoxyuridine triphosphate pyrophosphatase (DUT) suppresses incorporation of uracil into genomic DNA during replication. Thermostable DUTs from hyperthermophilic archaea such as Thermococcus pacificus enhance PCR amplification by preventing misincorporation of dUTP generated by spontaneous deamination of dCTP. However, it is necessary to elucidate whether DUTs do not cause dNTP imbalances during PCR by unwanted side activity. Moreover, it has been unknown what structural features define the thermostability of those DUTs. Here, DUT from a hyperthermophilic eubacterium, Aquifex aeolicus (Aa-DUT), was characterized together with those from T. pacificus (Tp-DUT). Aa-DUT was as thermostable as Tp-DUT up to at least 95°C. The crystal structures of the two thermostable enzymes were determined, which revealed that the structures of Aa-DUT and Tp-DUT resembled those of monofunctional and bifunctional DUTs, respectively. Generally, bifunctional DUTs harbor the dCTP deaminase activity in addition to the DUT activity. However, not only Aa-DUT but also Tp-DUT showed poor activity towards dCTP, indicating both enzymes are monofunctional. We further examined eight types of parameters related to thermostability of protein structure and found that the thermostability of Aa-DUT and Tp-DUT might be accomplished by increased numbers of ion pairs on the protein surface. Finally, we verified that Aa-DUT promoted PCR amplification with Pfu DNA polymerase to the same extent as Tp-DUT. Collectively, we conclude that both DUTs from hyperthermophiles maintain the enzymatic activity at high temperatures without consuming dCTP due to the lack of the deaminate activity., Nov. 2024, 33, 11, e5185, Scientific journal, True, 10.1002/pro.5185
- Refereed, Journal of Molecular Biology, Identification of a Catalytic Lysine Residue Conserved among GHKL ATPases: MutL, GyrB, and MORC., Kenji Fukui; Yuki Fujii; Takato Yano, DNA mismatch repair endonuclease MutL is a member of GHKL ATPase superfamily. Mutations of MutL homologs are causative of a hereditary cancer, Lynch syndrome. We characterized MutL homologs from human and a hyperthermophile, Aquifex aeolicus, (aqMutL) to reveal the catalytic mechanism for the ATPase activity. Although involvement of a basic residue had not been conceived in the catalytic mechanism, analysis of the pH dependence of the aqMutL ATPase activity revealed that the reaction is catalyzed by a residue with an alkaline pKa. Analyses of mutant aqMutLs showed that Lys79 is the catalytic residue, and the corresponding residues were confirmed to be critical for activities of human MutL homologs, on the basis of which a catalytic mechanism for MutL ATPase is proposed. These and other results described here would contribute to evaluating the pathogenicity of Lynch syndrome-associated missense mutations. Furthermore, it was confirmed that the catalytic lysine residue is conserved among DNA gyrases and microrchidia ATPases, other members of GHKL ATPases, indicating that the catalytic mechanism proposed here is applicable to these members of the superfamily., 17 Apr. 2024, 436, 10, 168575, 168575, Scientific journal, True, 10.1016/j.jmb.2024.168575
- Refereed, FEMS Microbiology Letters, An extensive ion-pair/hydrogen-bond network contributes to the thermostability of the MutL ATPase domain from Aquifex aeolicus., Ayaka Shibuya; Maki Yokote; Atsushi Suzuki; Kenji Fukui; Takato Yano, Proteins from hyperthermophiles often contain a large number of ionic interactions. Close examination of the previously determined crystal structure of the ATPase domain of MutL from a hyperthermophile, Aquifex aeolicus, revealed that the domain contains a continuous ion-pair/hydrogen-bond network consisting of 11 charged amino acid residues on a β-sheet. Mutations were introduced to disrupt the network, showing that the more extensively the network was disrupted, the greater the thermostability of the protein was decreased. Based on urea denaturation analysis, a thermodynamic parameter, energy for the conformational stability, was evaluated, which indicated that amino acid residues in the network contributed additively to the protein stability. A continuous network rather than a cluster of isolated interactions would pay less entropic penalty upon fixing the side chains to make the same number of ion pairs/hydrogen bonds, which might contribute more favorably to the structural formation of thermostable proteins., 21 Mar. 2024, 371, fnae020, Scientific journal, True, 10.1093/femsle/fnae020
- Not Refereed, Dec. 2023, 82, 2, 16, 24, Research institution
- Refereed, Scientific Reports, Springer Science and Business Media LLC, Characterization of cytoskeletal and structural effects of INF2 variants causing glomerulopathy and neuropathy, Hiroko Ueda; Quynh Thuy Huong Tran; Linh Nguyen Truc Tran; Koichiro Higasa; Yoshiki Ikeda; Naoyuki Kondo; Masaki Hashiyada; Chika Sato; Yoshinori Sato; Akira Ashida; Saori Nishio; Yasunori Iwata; Hiroyuki Iida; Daisuke Matsuoka; Yoshihiko Hidaka; Kenji Fukui; Suzu Itami; Norihito Kawashita; Keisuke Sugimoto; Kandai Nozu; Motoshi Hattori; Hiroyasu Tsukaguchi, Abstract
Focal segmental glomerulosclerosis (FSGS) is a common glomerular injury leading to end-stage renal disease. Monogenic FSGS is primarily ascribed to decreased podocyte integrity. Variants between residues 184 and 245 of INF2, an actin assembly factor, produce the monogenic FSGS phenotype. Meanwhile, variants between residues 57 and 184 cause a dual-faceted disease involving peripheral neurons and podocytes (Charcot–Marie–Tooth CMT/FSGS). To understand the molecular basis for INF2 disorders, we compared structural and cytoskeletal effects of INF2 variants classified into two subgroups: One (G73D, V108D) causes the CMT/FSGS phenotype, and the other (T161N, N202S) produces monogenic FSGS. Molecular dynamics analysis revealed that all INF2 variants show distinct flexibility compared to the wild-type INF2 and could affect stability of an intramolecular interaction between their N- and C-terminal segments. Immunocytochemistry of cells expressing INF2 variants showed fewer actin stress fibers, and disorganization of cytoplasmic microtubule arrays. Notably, CMT/FSGS variants caused more prominent changes in mitochondrial distribution and fragmentation than FSGS variants and these changes correlated with the severity of cytoskeletal disruption. Our results indicate that CMT/FSGS variants are associated with more severe global cellular defects caused by disrupted cytoskeleton-organelle interactions than are FSGS variants. Further study is needed to clarify tissue-specific pathways and/or cellular functions implicated in FSGS and CMT phenotypes, 25 Jul. 2023, 13, 1, Scientific journal, 10.1038/s41598-023-38588-7 - Refereed, Life Science Alliance, Life Science Alliance, LLC, Catalytic mechanism of the zinc-dependent MutL endonuclease reaction, Kenji Fukui; Tatsuya Yamamoto; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Takato Yano, DNA mismatch repair endonuclease MutL binds two zinc ions. However, the endonuclease activity of MutL is drastically enhanced by other divalent metals such as manganese, implying that MutL binds another catalytic metal at some site other than the zinc-binding sites. Here, we solved the crystal structure of the endonuclease domain ofAquifex aeolicusMutL in the manganese- or cadmium-bound form, revealing that these metals compete with zinc at the same sites. Mass spectrometry revealed that the MutL yielded 5′-phosphate and 3′-OH products, which is characteristic of the two-metal-ion mechanism. Crystallographic analyses also showed that the position and flexibility of a highly conserved Arg ofA. aeolicusMutL altered depending on the presence of zinc/manganese or the specific inhibitor cadmium. Site-directed mutagenesis revealed that the Arg was critical for the catalysis. We propose that zinc ion and its binding sites are physiologically of catalytic importance and that the two-metal-ion mechanism works in the reaction, where the Arg plays a catalytic role. Our results also provide a mechanistic insight into the inhibitory effect of a mutagen/carcinogen, cadmium, on MutL., 24 Jul. 2023, 6, 10, e202302001, e202302001, Scientific journal, 10.26508/lsa.202302001
- Refereed, Acta crystallographica. Section D, Structural biology, Serial femtosecond X-ray crystallography of an anaerobically formed catalytic intermediate of copper amine oxidase., Takeshi Murakawa; Mamoru Suzuki; Kenji Fukui; Tetsuya Masuda; Michihiro Sugahara; Kensuke Tono; Tomoyuki Tanaka; So Iwata; Eriko Nango; Takato Yano; Katsuyuki Tanizawa; Toshihide Okajima, The mechanisms by which enzymes promote catalytic reactions efficiently through their structural changes remain to be fully elucidated. Recent progress in serial femtosecond X-ray crystallography (SFX) using X-ray free-electron lasers (XFELs) has made it possible to address these issues. In particular, mix-and-inject serial crystallography (MISC) is promising for the direct observation of structural changes associated with ongoing enzymic reactions. In this study, SFX measurements using a liquid-jet system were performed on microcrystals of bacterial copper amine oxidase anaerobically premixed with a substrate amine solution. The structure determined at 1.94 Å resolution indicated that the peptidyl quinone cofactor is in equilibrium between the aminoresorcinol and semiquinone radical intermediates, which accumulate only under anaerobic single-turnover conditions. These results show that anaerobic conditions were well maintained throughout the liquid-jet SFX measurements, preventing the catalytic intermediates from reacting with dioxygen. These results also provide a necessary framework for performing time-resolved MISC to study enzymic reaction mechanisms under anaerobic conditions., 01 Dec. 2022, 78, Pt 12, 1428, 1438, Scientific journal, True, 10.1107/S2059798322010385
- Refereed, Journal of Structural Biology, Elsevier BV, Crystal structure of a nucleotide-binding domain of fatty acid kinase FakA from Thermus thermophilus HB8, Maya Nakatani; Shun-ya Nakahara; Kenji Fukui; Momoka Urano; Yuki Fujii; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Hiroki Okanishi; Yoshikatsu Kanai; Takato Yano; Ryoji Masui, Fatty acid kinase is necessary for the incorporation of exogenous fatty acids into membrane phospholipids. Fatty acid kinase consists of two components: a kinase component, FakA, that phosphorylates a fatty acid bound to a fatty acid-binding component, FakB. However, the molecular details underlying the phosphotransfer reaction remain to be resolved. We determined the crystal structure of the N-terminal domain of FakA bound to ADP from Thermus thermophilus HB8. The overall structure of this domain showed that the helical barrel fold is similar to the nucleotide-binding component of dihydroxyacetone kinase. The structure of the nucleotide-binding site revealed the roles of the conserved residues in recognition of ADP and Mg2+, but the N-terminal domain of FakA lacked the ADP-capping loop found in the dihydroxyacetone kinase component. Based on the structural similarity to the two subunits of dihydroxyacetone kinase complex, we constructed a model of the complex of T. thermophilus FakB and the N-terminal domain of FakA. In this model, the invariant Arg residue of FakB occupied a position that was spatially similar to that of the catalytically important Arg residue of dihydroxyacetone kinase, which predicted a composite active site in the Fatty acid kinase complex., Dec. 2022, 214, 4, 107904, 107904, Scientific journal, True, 10.1016/j.jsb.2022.107904
- Refereed, Structure, Structural and functional insights into the mechanism by which MutS2 recognizes a DNA junction, Kenji Fukui; Masao Inoue; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Takato Yano, Apr. 2022, 30, 7, 973, 982, Scientific journal
- Refereed, Acta Crystallographica Section F Structural Biology Communications, International Union of Crystallography (IUCr), Microcrystal preparation for serial femtosecond X-ray crystallography of bacterial copper amine oxidase, Takeshi Murakawa; Mamoru Suzuki; Toshi Arima; Michihiro Sugahara; Tomoyuki Tanaka; Rie Tanaka; So Iwata; Eriko Nango; Kensuke Tono; Hideyuki Hayashi; Kenji Fukui; Takato Yano; Katsuyuki Tanizawa; Toshihide Okajima, Recent advances in serial femtosecond X-ray crystallography (SFX) using X-ray free-electron lasers have paved the way for determining radiation-damage-free protein structures under nonfreezing conditions. However, the large-scale preparation of high-quality microcrystals of uniform size is a prerequisite for SFX, and this has been a barrier to its widespread application. Here, a convenient method for preparing high-quality microcrystals of a bacterial quinoprotein enzyme, copper amine oxidase from Arthrobacter globiformis, is reported. The method consists of the mechanical crushing of large crystals (5–15 mm3), seeding the crushed crystals into the enzyme solution and standing for 1 h at an ambient temperature of ∼26°C, leading to the rapid formation of microcrystals with a uniform size of 3–5 µm. The microcrystals diffracted X-rays to a resolution beyond 2.0 Å in SFX measurements at the SPring-8 Angstrom Compact Free Electron Laser facility. The damage-free structure determined at 2.2 Å resolution was essentially identical to that determined previously by cryogenic crystallography using synchrotron X-ray radiation., 01 Oct. 2021, 77, 10, 356, 363, Scientific journal, True, 10.1107/s2053230x21008967
- Refereed, Journal of Biological Chemistry, A Lynch syndrome-associated mutation at a Bergerat ATP-binding fold destabilizes the structure of the DNA mismatch repair endonuclease MutL, Keisuke Izuhara; Kenji Fukui; Takeshi Murakawa; Seiki Baba; Takashi Kumasaka; Kazuhisa Uchiyama; Takato Yano, In humans, mutations in genes encoding homologs of the DNA mismatch repair endonuclease MutL cause a hereditary cancer that is known as Lynch syndrome. Here, we determined the crystal structures of the N-terminal domain (NTD) of MutL from the thermophilic eubacterium Aquifex aeolicus (aqMutL) complexed with ATP analogs at 1.69-1.73 Å. The structures revealed significant structural similarities to those of a human MutL homolog, postmeiotic segregation increased 2 (PMS2). We introduced five Lynch syndrome-associated mutations clinically found in human PMS2 into the aqMutL NTD and investigated the protein stability, ATPase activity, and DNA-binding ability of these protein variants. Among the mutations studied, the most unexpected results were obtained for the residue Ser34. Ser34 (Ser46 in PMS2) is located at a previously identified Bergerat ATP-binding fold. We found that the S34I aqMutL NTD retains ATPase and DNA-binding activities. Interestingly, CD spectrometry and trypsin-limited proteolysis indicated the disruption of a secondary structure element of the S34I NTD, destabilizing the overall structure of the aqMutL NTD. In agreement with this, the recombinant human PMS2 S46I NTD was easily digested in the host Escherichia coli cells. Moreover, other mutations resulted in reduced DNA-binding or ATPase activity. In summary, using the thermostable aqMutL protein as a model molecule, we have experimentally determined the effects of the mutations on MutL endonuclease; we discuss the pathological effects of the corresponding mutations in human PMS2., Aug. 2020, 295, 33, 11643, 11655, Scientific journal, True, False, 10.1074/jbc.RA120.013576
- Refereed, Enzyme and Microbial Technology, Catalytic properties and crystal structure of UDP-galactose 4-epimerase-like l-threonine 3-dehydrogenase from Phytophthora infestans, Kazunari Yoneda; Rina Nagano; Takuya Mikami; Haruhiko Sakuraba; Kenji Fukui; Tomohiro Araki; Toshihisa Ohshima, Jul. 2020, 140, 109627
- Refereed, DNA Repair, Biochemical characterization of mismatch-binding protein MutS1 and nicking endonuclease MutL from a euryarchaeon Methanosaeta thermophila., Minobe A; Fukui K; Yonezu H; Ohshita K; Mizobuchi S; Morisawa T; Hakumai Y; Yano T; Ashiuchi M; Wakamatsu T, Jan. 2019, 75, 29, 38, Scientific journal
- Refereed, FEBS Letters, The GIY‐YIG endonuclease domain of Arabidopsis MutS homolog 1 specifically binds to branched DNA structures, Kenji Fukui; Akiko Harada; Taisuke Wakamatsu; Ai Minobe; Koki Ohshita; Makoto Ashiuchi; Takato Yano, Oct. 2018, 592, 24, 4066, 4077, Scientific journal
- Refereed, J. Bacteriol., Resistance to UV Irradiation Caused by Inactivation of nurA and herA Genes in Thermus thermophilus., Yuki Fujii; Masao Inoue; Kenji Fukui; Seiki Kuramitsu; Ryoji Masui, Jul. 2018, 200, 16, e00201, e00218, Scientific journal
- Refereed, FEBS Letters, Wiley Blackwell, Multiple zinc ions maintain the open conformation of the catalytic site in the DNA mismatch repair endonuclease MutL from Aquifex aeolicus, Kenji Fukui; Seiki Baba; Takashi Kumasaka; Takato Yano, 01 May 2018, 592, 9, 1611, 1619, Scientific journal, 10.1002/1873-3468.13050
- Refereed, CHEMICAL COMMUNICATIONS, A covalent G-site inhibitor for glutathione S-transferase Pi (GSTP(1-1)), Yuko Shishido; Fumiaki Tomoike; Yasuaki Kimura; Keiko Kuwata; Takato Yano; Kenji Fukui; Haruka Fujikawa; Yoshitaka Sekido; Yuko Murakami-Tonami; Tomoshi Kameda; Satoshi Shuto; Hiroshi Abe, Oct. 2017, 53, 81, 11138, 11141, Scientific journal, 10.1039/c7cc05829b
- Refereed, FEBS JOURNAL, Archaeal MutS5 tightly binds to Holliday junction similarly to eukaryotic MutS gamma, Koki Ohshita; Kenji Fukui; Mizuki Sato; Takashi Morisawa; Yuichi Hakumai; Yuki Morono; Fumio Inagaki; Takato Yano; Makoto Ashiuchi; Taisuke Wakamatsu, Oct. 2017, 284, 20, 3470, 3483, Scientific journal, 10.1111/febs.14204
- Refereed, Biochemistry and Biophysics Reports, Elsevier B.V., Indispensable residue for uridine binding in the uridine-cytidine kinase family, Fumiaki Tomoike; Noriko Nakagawa; Kenji Fukui; Takato Yano; Seiki Kuramitsu; Ryoji Masui, 01 Sep. 2017, 11, 93, 98, Scientific journal, 10.1016/j.bbrep.2017.07.002
- Refereed, Biochim Biophys Acta, Crystal structure and DNA-binding property of the ATPase domain of bacterial mismatch repair endonuclease MutL from Aquifex aeolicus, Kenji Fukui; Hitoshi Iino; Seiki Baba; Takashi Kumasaka; Seiki Kuramitsu; Takato Yano, Jun. 2017, 1865, 9, 1178, 1187, Scientific journal
- Refereed, SCIENTIFIC REPORTS, High-throughput Screening of Small Molecule Inhibitors of the Streptococcus Quorum-sensing Signal Pathway, Seiji Ishii; Kenji Fukui; Satoshi Yokoshima; Kazuo Kumagai; Youko Beniyama; Tetsuya Kodama; Tohru Fukuyama; Takayoshi Okabe; Tetsuo Nagano; Hirotatsu Kojima; Takato Yano, Jun. 2017, 7, 1, 4029, Scientific journal, 10.1038/s41598-017-03567-2
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, The Lon protease-like domain in the bacterial RecA paralog RadA is required for DNA binding and repair, Masao Inoue; Kenji Fukui; Yuki Fujii; Noriko Nakagawa; Takato Yano; Seiki Kuramitsu; Ryoji Masui, Jun. 2017, 292, 23, 9801, 9814, Scientific journal, 10.1074/jbc.M116.770180
- Refereed, Biochimica et Biophysica Acta - Proteins and Proteomics, Elsevier B.V., Proteome-wide identification of lysine succinylation in thermophilic and mesophilic bacteria, Hiroki Okanishi; Kwang Kim; Kenji Fukui; Takato Yano; Seiki Kuramitsu; Ryoji Masui, 01 Feb. 2017, 1865, 2, 232, 242, Scientific journal, 10.1016/j.bbapap.2016.11.009
- Refereed, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Structural Insights into L-Tryptophan Dehydrogenase from a Photoautotrophic Cyanobacterium, Nostoc punctiforme, Taisuke Wakamatsu; Haruhiko Sakuraba; Megumi Kitamura; Yuichi Hakumai; Kenji Fukui; Kouhei Ohnishi; Makoto Ashiuchi; Toshihisa Ohshima, Jan. 2017, 83, 2, e02710, e02716, Scientific journal, 10.1128/AEM.02710-16
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, Structural Features and Functional Dependency on -Clamp Define Distinct Subfamilies of Bacterial Mismatch Repair Endonuclease MutL, Kenji Fukui; Seiki Baba; Takashi Kumasaka; Takato Yano, Aug. 2016, 291, 33, 16990, 17000, Scientific journal, 10.1074/jbc.M116.739664
- Refereed, EXTREMOPHILES, Roles of Mn-catalase and a possible heme peroxidase homologue in protection from oxidative stress in Thermus thermophilus, Akio Ebihara; Miho Manzoku; Kenji Fukui; Atsuhiro Shimada; Rihito Morita; Ryoji Masui; Seiki Kuramitsu, Jul. 2015, 19, 4, 775, 785, Scientific journal, 10.1007/s00792-015-0753-2
- Refereed, EXTREMOPHILES, Small-angle X-ray scattering analysis reveals the ATP-bound monomeric state of the ATPase domain from the homodimeric MutL endonuclease, a GHKL phosphotransferase superfamily protein, Hitoshi Iino; Takaaki Hikima; Yuya Nishida; Masaki Yamamoto; Seiki Kuramitsu; Kenji Fukui, May 2015, 19, 3, 643, 656, Scientific journal, 10.1007/s00792-015-0745-2
- Refereed, PLOS ONE, NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP, Ryota Mizushima; Ju Yaen Kim; Isao Suetake; Hiroaki Tanaka; Tomoyo Takai; Narutoshi Kamiya; Yu Takano; Yuichi Mishima; Shoji Tajima; Yuji Goto; Kenji Fukui; Young-Ho Lee, Jun. 2014, 9, 6, e98554, k.fukui, Scientific journal, 10.1371/journal.pone.0098554
- Refereed, BMC GENOMICS, The role of ribonucleases in regulating global mRNA levels in the model organism Thermus thermophilus HB8, Hiromasa Ohyama; Tomofumi Sakai; Yoshihiro Agari; Kenji Fukui; Noriko Nakagawa; Akeo Shinkai; Ryoji Masui; Seiki Kuramitsu, May 2014, 15, 386, Scientific journal, 10.1186/1471-2164-15-386
- Refereed, FEBS JOURNAL, MutS stimulates the endonuclease activity of MutL in an ATP-hydrolysis-dependent manner, Atsuhiro Shimada; Yoshitaka Kawasoe; Yoshito Hata; Tatsuro S. Takahashi; Ryoji Masui; Seiki Kuramitsu; Kenji Fukui, Jul. 2013, 280, 14, 3467, 3479, Scientific journal, 10.1111/febs.12344
- Refereed, INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR), Kenji Fukui; Yoshitaka Bessho; Atsuhiro Shimada; Shigeyuki Yokoyama; Seiki Kuramitsu, Mar. 2013, 14, 3, 6436, 6453, Scientific journal, 10.3390/ijms14036436
- Refereed, Journal of Nucleic Acids, Hindawi Limited, Simultaneous Use of MutS and RecA for Suppression of Nonspecific Amplification during PCR, Kenji Fukui; Seiki Kuramitsu, Thermus thermophilusMutS, a thermostable mismatch-recognizing protein, is utilized in PCR to suppress nonspecific amplification by preventing synthesis from mismatched primers.T. thermophilusRecA also decreases nonspecific amplification by promoting proper hybridization between the primer and template. We observed that MutS and RecA function under the same reaction conditions and that MutS and RecA do not preclude each other. Furthermore, there were some DNA sequences for which only one of the 2 proteins effectively suppressed nonspecific amplification. The simultaneous use of MutS and RecA is a more attractive error-suppressing technique than the use of either of the 2 proteins alone., 2013, 2013, 1, 5, Scientific journal, 10.1155/2013/823730
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, Evidence for ATP-dependent Structural Rearrangement of Nuclease Catalytic Site in DNA Mismatch Repair Endonuclease MutL, Tatsuya Yamamoto; Hitoshi Iino; Kwang Kim; Seiki Kuramitsu; Kenji Fukui, Dec. 2011, 286, 49, 42337, 42348, Scientific journal, 10.1074/jbc.M111.277335
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, In Vivo, in Vitro, and X-ray Crystallographic Analyses Suggest the Involvement of an Uncharacterized Triose-phosphate Isomerase (TIM) Barrel Protein in Protection against Oxidative Stress, Shuhei Nakane; Taisuke Wakamatsu; Ryoji Masui; Seiki Kuramitsu; Kenji Fukui, Dec. 2011, 286, 48, 41636, 41646, Scientific journal, 10.1074/jbc.M111.293886
- Refereed, Bioscience Reports, Characterization of C- and N-terminal domains of Aquifex aeolicus MutL endonuclease: N-terminal domain stimulates the endonuclease activity of C-terminal domain in a zinc-dependent manner, Hitoshi Iino; Kwang Kim; Atsuhiro Shimada; Ryoji Masui; Seiki Kuramitsu; Kenji Fukui, Oct. 2011, 31, 5, 309, 322, Scientific journal, 10.1042/BSR20100116
- Refereed, JOURNAL OF BIOCHEMISTRY, An alkyltransferase-like protein from Thermus thermophilus HB8 affects the regulation of gene expression in alkylation response, Rihito Morita; Hisahiro Hishinuma; Hiromasa Ohyama; Ryosuke Mega; Toshihiro Ohta; Noriko Nakagawa; Yoshihiro Agari; Kenji Fukui; Akeo Shinkai; Seiki Kuramitsu; Ryoji Masui, Sep. 2011, 150, 3, 327, 339, Scientific journal, 10.1093/jb/mvr052
- Refereed, JOURNAL OF BIOCHEMISTRY, Crystal structure of the tandem-type universal stress protein TTHA0350 from Thermus thermophilus HB8, Hitoshi Iino; Nobutaka Shimizu; Masaru Goto; Akio Ebihara; Kenji Fukui; Ken Hirotsu; Seiki Kuramitsu, Sep. 2011, 150, 3, 295, 302, Scientific journal, 10.1093/jb/mvr057
- Refereed, PLOS ONE, Inactivation of the DNA Repair Genes mutS, mutL or the Anti-Recombination Gene mutS2 Leads to Activation of Vitamin B-1 Biosynthesis Genes, Kenji Fukui; Taisuke Wakamatsu; Yoshihiro Agari; Ryoji Masui; Seiki Kuramitsu, Apr. 2011, 6, 4, e19053, Scientific journal, 10.1371/journal.pone.0019053
- Journal of nucleic acids, Molecular mechanisms of the whole DNA repair system: a comparison of bacterial and eukaryotic systems., Rihito Morita; Shuhei Nakane; Atsuhiro Shimada; Masao Inoue; Hitoshi Iino; Taisuke Wakamatsu; Kenji Fukui; Noriko Nakagawa; Ryoji Masui; Seiki Kuramitsu, DNA is subjected to many endogenous and exogenous damages. All organisms have developed a complex network of DNA repair mechanisms. A variety of different DNA repair pathways have been reported: direct reversal, base excision repair, nucleotide excision repair, mismatch repair, and recombination repair pathways. Recent studies of the fundamental mechanisms for DNA repair processes have revealed a complexity beyond that initially expected, with inter- and intrapathway complementation as well as functional interactions between proteins involved in repair pathways. In this paper we give a broad overview of the whole DNA repair system and focus on the molecular basis of the repair machineries, particularly in Thermus thermophilus HB8., 14 Oct. 2010, 2010, 179594, 179594, Scientific journal, True, 10.4061/2010/179594
- Refereed, NUCLEIC ACIDS RESEARCH, A novel single-stranded DNA-specific 3'-5' exonuclease, Thermus thermophilus exonuclease I, is involved in several DNA repair pathways, Atsuhiro Shimada; Ryoji Masui; Noriko Nakagawa; Yoshio Takahata; Kwang Kim; Seiki Kuramitsu; Kenji Fukui, Sep. 2010, 38, 17, 5692, 5705, Scientific journal, 10.1093/nar/gkq350
- Refereed, Journal of Nucleic Acids, Hindawi Limited, DNA Mismatch Repair in Eukaryotes and Bacteria, Kenji Fukui, DNA mismatch repair (MMR) corrects mismatched base pairs mainly caused by DNA replication errors. The fundamental mechanisms and proteins involved in the early reactions of MMR are highly conserved in almost all organisms ranging from bacteria to human. The significance of this repair system is also indicated by the fact that defects in MMR cause human hereditary nonpolyposis colon cancers as well as sporadic tumors. To date, 2 types of MMRs are known: the human type andEscherichia colitype. The basic features of the former system are expected to be universal among the vast majority of organisms including most bacteria. Here, I review the molecular mechanisms of eukaryotic and bacterial MMR, emphasizing on the similarities between them., 2010, 2010, 1, 16, Scientific journal, 10.4061/2010/260512
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, Crystal Structure of MutS2 Endonuclease Domain and the Mechanism of Homologous Recombination Suppression, Kenji Fukui; Noriko Nakagawa; Yoshiaki Kitamura; Yuya Nishida; Ryoji Masui; Seiki Kuramitsu, Nov. 2008, 283, 48, 33417, 33427, Scientific journal, 10.1074/jbc.M806755200
- Refereed, JOURNAL OF BIOLOGICAL CHEMISTRY, Bound nucleotide controls the endonuclease activity of mismatch repair enzyme MutL, Kenji Fukui; Masami Nishida; Noriko Nakagawa; Ryoji Masui; Seiki Kuramitsu, May 2008, 283, 18, 12136, 12145, Scientific journal, 10.1074/jbc.M800110200
- Refereed, Nucleic Acids Research, 15, Analysis of a nuclease activity of catalytic domain of Thermus thermophilus MutS2 by high-accuracy mass spectrometry, Kenji Fukui; Yoshio Takahata; Noriko Nakagawa; Seiki Kuramitsu; Ryoji Masui, Aug. 2007, 35, 15, e100, k.fukui, Scientific journal, 10.1093/nar/gkm575
- Refereed, NATURE METHODS, Rapid SNP diagnostics using asymmetric isothermal amplification and a new mismatch-suppression technology, Yasumasa Mitani; Alexander Lezhava; Yuki Kawai; Takeshi Kikuchi; Atsuko Oguchi-Katayama; Yasushi Kogo; Masayoshi Itoh; Toru Miyagi; Hideki Takakura; Kanako Hoshi; Chiaki Kato; Takahiro Arakawa; Kazuhiro Shibata; Kenji Fukui; Ryoji Masui; Seiki Kuramitsu; Kazuma Kiyotani; Alistair Chalk; Katsuhiko Tsunekawa; Masami Murakami; Tetsuya Kamataki; Takanori Oka; Hiroshi Shimada; Paul E. Cizdziel; Yoshihide Hayashizaki, Mar. 2007, 4, 3, 257, 262, Scientific journal, 10.1038/NMETH1007
- Refereed, NUCLEIC ACIDS RESEARCH, Nuclease activity of the MutS homologue MutS2 from Thermus thermophilus is confined to the Smr domain, Kenji Fukui; Hiromichi Kosaka; Seiki Kuramitsu; Ryoji Masui, Feb. 2007, 35, 3, 850, 860, Scientific journal, 10.1093/nar/grl735
- Refereed, JOURNAL OF BIOCHEMISTRY, Thermus thermophilus MutS2, a MutS paralogue, possesses an endonuclease activity promoted by MutL, K Fukui; R Masui; S Kuramitsu, Mar. 2004, 135, 3, 375, 384, Scientific journal, 10.1093/jb/mvh045
MISC
- 2024, 68th
- 日本生化学会大会プログラム・講演要旨集, (公社)日本生化学会, TRPC6遺伝子変異による糸球体障害に関する分子遺伝学的研究(Molecular Genetics Underlying Glomerulopathy Caused by TRPC6 Mutations), トラン・チュイフォンクィン; 福井 健二; 塚口 裕康, Oct. 2023, 96回, [1P, 351]
- 2023, 2023 (CD-ROM)
- Nov. 2021, 94回, [1T15m, 208)]
- Nov. 2021, 94回, [P, 227]
- Sep. 2021, 80, 1・2, 85, 91
- 2020, 56th
- 2020, 93rd
- 2020, 43rd
- Mar. 2019, 139年会, 2, 80, 80
- 2019, 53rd
- 2019, 53rd
- 2019, 42nd
- 2019, 71st
- 2019, 53rd
- ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, Development of glutathione S-transferase (GST) covalent inhibitor, Yuko Shishido; Fumiaki Tomoike; Yasuaki Kimura; Keiko Kuwata; Takato Yano; Kenji Fukui; Haruka Fujikawa; Yoshitaka Sekido; Yuko Murakami-Tonami; Tomoshi Kameda; Satoshi Shuto; Hiroshi Abe, Mar. 2018, 255, Summary international conference
- 2018, 91st
- Not Refereed, Dec. 2017, 2017年度, [3P, 0205]
- Dec. 2017, 2017年度, [1AW10, 4]
- Dec. 2017, 2017年度, [3P, 0205]
- Dec. 2017, 2017年度, [3LBA, 007]
- Dec. 2017, 2017年度, [4AT26, 1420)]
- Mar. 2017, 137年会, 2, 68, 68
- 2017, 17th
- 2017, 48th
- 2016, 16th
- Dec. 2015, 88回・38回, [2P0681], [2P0681]
- Dec. 2015, 88回・38回, [2P0683], [2P0683]
- 2015, 54th
- Refereed, Seikagaku, Japanese Biochemical Society, Regulatory mechanism for DNA mismatch repair endonuclease, Kenji Fukui, 2015, 87, 2, 212, 217, Introduction scientific journal, 10.14952/SEIKAGAKU.2015.870212
- Refereed, Vitamins (Japan), Inactivation of the DNA repair genes mutS, mutL or the anti-recombination gene mutS2 leads to activation of vitamin B1 biosynthesis genes, Kenji Fukui; Taisuke Wakamatsu; Yoshihiro Agari; Ryoji Masui; Seiki Kuramitsu, Sep. 2014, 88, 9, 459, 461, Introduction other, 10.20632/vso.88.9_459
- Sep. 2013, 86回, 3P, 471
- 2013, 7th
- 2013, 13th
- 2013, 61st
- 2013, 36th
- 2013, 36th
- 2013, 36th
- Dec. 2012, 85回, 2P, 167
- Dec. 2012, 85回, 2P, 610
- Dec. 2012, 85回, 2P, 615
- Dec. 2012, 85回, 2P, 617
- 2012, 51st
- 2012, 85th
- 2012, 35th
- 2012, 85th
- Refereed, DNA Repair - on the pathways to fixing DNA damage and errors, Biochemical Properties of MutL, a DNA Mismatch Repair Endonuclease, Kenji Fukui; Atsuhiro Shimada; Hitoshi Iino; Ryoji Masui; Seiki Kuramitsu, Sep. 2011, 123, 142, Introduction scientific journal
- 2011, 161st
- 2011, 11th
- 2010
- 2010, 10th
- 2010
- 2010
- 2010, 10th
- 2010, 10th
- 2010, 10th
- 2010
- Sep. 2009, 82回, 3P, 574
- 2009
- Not Refereed, Acta Crystallographica Section A Foundations of Crystallography, International Union of Crystallography (IUCr), Structural and functional whole-cell project for the model organism, Thermus thermophilus HB8, Akio Ebihara; Akeo Shinkai; Mayumi Kanagawa; Yoshihiro Agari; Hitoshi Iino; Yoshiaki Kitamura; Keiko Sakamoto; Miho Manzoku; Kenji Fukui; Noriko Nakagawa; Ryoji Masui; Ken Hirotsu; Yoshitaka Bessho; Takaho Terada; Mikako Shirouzu; Shigeyuki Yokoyama, Aug. 2008, 64, a1, C361, Summary international conference, 10.1107/s0108767308088466
- 2008, 8th
- 2008
- 2008
- 2007
- 2007
- 2007
- 2006, 6th
- 2005, 28th
- 2005, 28th
- 2002, 74, 8
Awards
Research Projects
- Aug. 2025 - Mar. 2028, Principal investigator, MutS ファミリータンパク質の進化ダイナミクス： DNA 修復と組換えのための構造・機能の多様化, 福井健二, 山田科学振興財団, 2025年度研究援助_チャレンジ支援枠, 奈良女子大学, url
- Grant-in-Aid for Scientific Research (C), 01 Apr. 2024 - 31 Mar. 2027, 24K08718, High-Temperature X-ray Crystallography on the Proteins from Thermophiles, Opening the Way to Protein Science in Extreme Environments, 福井 健二; 馬場 清喜; 村川 武志, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Osaka Medical and Pharmaceutical University, 4420000, 3400000, 1020000, kaken
- 挑戦的研究(萌芽), 30 Jun. 2023 - 31 Mar. 2026, 23K18117, 速度論および熱力学的手法を用いた"in crystallo" 酵素反応解析, 村川 武志; 馬場 清喜; 福井 健二, 日本学術振興会, 科学研究費助成事業, 大阪医科薬科大学, 6500000, 5000000, 1500000, kaken
- Apr. 2023 - Mar. 2025, G-2023-2-048, Principal investigator, (none), 福井健二, Institute for Fermentation, Osaka, 一般研究助成, 3000000, 3000000, Competitive research funding, url
- Grant-in-Aid for Scientific Research (C), Apr. 2022 - Mar. 2025, 22K09953, Coinvestigator, (none), 矢野 貴人, 石井 誠志, 福井 健二, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Osaka Medical and Pharmaceutical University, 4160000, 3200000, 960000, kaken
- Grant-in-Aid for Scientific Research (C), Apr. 2019 - Mar. 2022, 19K10082, Coinvestigator, (none), YANO Takato, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Osaka Medical College, 4160000, 3200000, 960000, The genus Streptococcus is a member of commensal bacteria in oral cavity. However, once in the blood stream, it causes chronic infection by the formation of biofilm, which is resistant to treatments, such as antibiotics. The signal molecule for the initiation of biofilm formation in Streptococcus is generated and exported by a membrane protein, ComA. ComA is a multidomain protein comprising peptidase, nucleotide-binding, and transmembrane domains.
The full-length ComA protein was synthesized by wheat-germ in vitro expression system, and the protein could be partially purified by repeated liposome precipitation. The full-length ComA protein thus obtained was confirmed to be enzymatically active, and the cooperation between the peptidase and ATPase activities was analyzed. Further, the IC50 values for the full-length ComA and the peptidase domain and the EC50 value for the biofilm formation of S. mutans were compared., kaken - Apr. 2019 - Mar. 2022, Principal investigator, リンチ症候群発症機序の解明と創薬に向けたDNAミスマッチ修復蛋白質の構造機能解析, 福井 健二, 文部科学省, 科学研究費補助金(基盤研究(C)), 2990000, 2300000, 690000, Competitive research funding, url
- Grant-in-Aid for Scientific Research (C), 01 Apr. 2018 - 31 Mar. 2021, 18K06190, Mismatch repair system in prokaryotes lacking canonical mutS-dependent system, Takemoto Norihiko, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, National Center for Global Health and Medicine, 4290000, 3300000, 990000, MutS-dependent mismatch repair system have been known to be conserved from bacteria to human, as a replication error correction system. Disruption of MMR system results in increased mutation rate, leading to high occurrence of cancer cells or drug-resistant pathogens. However, recent advances in Next Generation Sequencing technologies shed light on the existence of exceptions, in which mutation rate is not so high regardless of the lack of canonical MMR system. In this study, we searched for conserved gene which works to decrease mutations in prokaryotes lacking canonical MMR system. We found that EndoMS works for replication error correction., kaken
- Apr. 2016 - Mar. 2019, Principal investigator, DNA/RNA ミスマッチ認識蛋白質を用いた高精度逆転写による相同 miRNA の識別, Kenji Fukui, Ministry of Education, Culture, Sports, Science and Technology, Grant-in-Aid for Encouragement of Young Scientists (B), 2470000, 1900000, 570000, Competitive research funding, url
- Grant-in-Aid for Young Scientists (B), Apr. 2014 - Mar. 2016, 26850050, Principal investigator, Suppression of errors in reverse transcription, Fukui Kenji, Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Osaka Medical College, 2990000, 2300000, 690000, Reverse transcription is essential for gene expression analysis, detection of pathogenic microorganisms, and other diagnostic technologies. In this study, we developed a novel method for suppressing non-specific amplifications during reverse transcription-related reactions. The previously-uncharacterized protein LLBP was isolated from Thermus thermophlus HB8 and its binding specificity towards DNA/RNA mismatch was confirmed. LLBP inhibited reverse transcription reaction when the primer-template complex contained a DNA/RNA mismatch. This protein would be utilized for improving the accuracy of reverse transcription-related technologies., Competitive research funding, url
- 若手研究(B), Apr. 2013 - Mar. 2015, 25850062, Principal investigator, 逆転写反応における非特異的増幅の抑制, 福井 健二, 文部科学省, 科学研究費補助金(若手研究(B)), 独立行政法人理化学研究所, 3250000, 2500000, 750000, Competitive research funding, url
- 若手研究(スタートアップ), Apr. 2008 - Mar. 2010, 20870042, Principal investigator, Structural and functional analyses on a novel mechanism of homologous recombination suppression., Kenji FUKUI, Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(若手研究(スタートアップ)), 独立行政法人理化学研究所, 2834000, 2180000, 654000, Recent studies revealed the existence of a novel anti-recombination enzyme, MutS2 ; however the mechanism by which MutS2 inhibits homologous recombination has been unknown. To elucidate the mechanism, we performed structural and functional analyses on Thermus thermophilus MutS2. X-ray crystallographic analysis on the C-terminal domain of MutS2 revealed that this domain resembles the structures of known endonucleases. Furthermore, biochemical experiments demonstrated that MutS2 preferentially incises the early intermediates in homologous recombination. These results indicate that MutS2 suppresses homologous recombination through a novel mechanism involving resolution of early intermediates., Competitive research funding, url
- 特別研究員奨励費, Apr. 2005 - Mar. 2007, 05J09716, Principal investigator, DNA修複において重要な役割をもつ、ミスマッチ修複系酵素群の構造機能解析, 福井 健二, 文部科学省, 科学研究費補助金(特別研究員奨励費), 大阪大学, 0, 0, 0, DNA修復系酵素MutSファミリーのうち、MutS2に着目し、その分子機能解析をおこなった。高度好熱菌mutS2の遺伝子破壊株を作製し、表現型を野生株と比較したところ、mutS2欠損株では、高い相同組換え効率を示すこと、マイトマイシンCに対する耐性が上昇していることが分かった。また、生化学的な解析から、MutS2は、相同組換えの一般的な中間体であるD-loop構造を認識し、切断する活性を持つ事が示唆された。さらに、D-loop構造を切断するヌクレアーゼ活性がMutS2のC末端に存在するSmrドメインに由来することを部位特異的変異導入法により明らかにした。このドメインはバクテリアからヒトまで高度に保存された機能未知のドメインであったが、本研究により新奇のヌクレアーゼドメインであることが示された。そこで、このドメインのヌクレアーゼ活性を詳細に調べるために、フーリエ変換型質量分析計を用いた解析法を確立した。この解析により、Smrドメインのヌクレアーゼ活性は、配列特異性を持たない事、ホスホジエステル結合をリン酸基の5'側で加水分解する事などが分かった。これは、フーリエ変換型質量分析計によりヌクレアーゼ活性を解析した初めての例である。以上の結果から、MutS2は相同組換え申間体を認識し、解離させることにより相同組換えを抑制すると考えられる。相同組換えは、外来DNAの取り込みなどを伸介する反応であり、これを制御することによりゲノムの安定性の維持に関与しているといえる。, Competitive research funding, url
