Laboratory of Molecular and Cellular Biology of Totipotency, Division of Integrated Life Science,

Graduate School of Biostudies, Kyoto University 　(updated partially, May 18, 2011)

(formerly, Laboratory of Molecular and Cellular Biology,Division of Applied Life Sciences,

Graduate School of Agriculture, Kyoto University)

<Japanese version here>

TEL +81-75-753-6381 / FAX +81-75-753-6398

Staff

Professor: Sato, Fumihiko, Dr. Agric. Sci.

Associate Professor: Endo, Tsuyoshi, Dr. Agric. Sci.

Instructor: Ifuku, Kentaro, Dr. Agric. Sci.

Students and research fellows

Post-doctoral fellows : (1), Doctor's program : (4), Master's program : (9), Undergraduate : (3)

A. Research Activities

a) Studies on functional differentiation of plant cells

　　　Cultured green cells which can grow without any addition of organic carbon sources have been established. Using these photoautotrophic cultured cells, the photosynthetic functions of plant have been investigated on cellular and molecular levels as follows. 1) Chracterization of regulation of gene expression in plant cells; e.g. cDNA of a chloroplast-DNA binding protein of 41kD (CND41) has been isolated and the accumlation of CND41 protein has been determined. A possible function of CND41 as a negative regulator of chloroplast gene expression has been studied. The regulatory mechanism of accumulation of neutral pathogenesis-related protein 5d (osmotin-like protein)in cultured tobacco cells has also characterized. The sequence analysis and studies with transgenic tobacco indicated that AGCCGCC sequence(s) in promoter region of OLP gene is a DNA element(s) responsive to ethylene and also to constitutive expression in root and cultured cells. 2) Selection of environmental stress-tolerant cells (plants) and their biochemical and molecular biological characterization. Using a cell line of tobacco that was able to grow photoautotrophically in a medium that contained 0.2 M NaCl, the salt tolerance mechanism in thylakoid membranes has been characterized. Especially, the functional role of 23-kDa polypeptides (OEC23) from the water-splitting complex of PSII has been studeied with in vitro reconstitution experiment using recombinant OEC23 and with transgenic plants. 3) Biochemical and molecular characterization of carbon fixation, especially on phosphoenolpyruvate carboxylase of C3 plants.

b) Molecular biology of secondary metabolism in plant cells

High metabolite-producing cells/tissues were established by the cellular selection or by the organ culture techniques. 1) Molecular characterization of isoquinoline alkaloid biosynthesis. Using high berberine-producing Coptis cells, major enzymes (O-methyltransferases, cytochrome P450 etc.) have been isolated, purified and characterized. cDNAs of these enzymes have also been isolated and molecular functions of enzymes and regulatory mechanism of their gene expression have been characterized. 2) Transport mechanism of the isoquinoline alkaloid and naphthoquinone derivatives in plant cells has been also investigated using berberine producing cell cultures, and shikonin producing cell cultures, respectively.

c) Molecular characterization of plant functions using transgenic plants

Using the genes isolated in our laboratory, transgenic plants which have altered expression of targeted gene were produced and the function(s) of gene(s) have been investigated as follows. 1) Molecular breeding of medicinal plants, e.g. metabolic engineering of isoquinoline alkaloid biosynthesis in Coptis cells, 2) Genetic manipulation of plant metabolism and stress-tolerance, using PEPCase cDNA, OLP cDNA and compatible solute-related genes, 4) Metabolic engineering of terpenoids in Lithospermum hairy root, and so on. To characterize the antifungal activity of OLP, OLP was crystallized and its three dimensional structure was resolved.

A-2. Publications and presentations

a) Publication (last 7 years; more Publications, please see (1994-))

Some selected original papers

Nakagawa A, Minami H, Kim JS, Koyanagi T, Katayama T, Sato F, Kumagai H. (2011) A bacterial platform for fermentative production of plant alkaloids. Nature Communications 2, Article number: 326, doi:10.1038/ncomms1327, Published 24 May 2011

Yasuyuki Yamada; Yasuhisa Kokabu; Kaori Chaki; Tadashi Yoshimoto; Mai Ohgaki; Sayumi Yoshida; Nobuhiko Kato; Tomotsugu Koyama; Fumihiko Sato　(2011) Isoquinoline alkaloid biosynthesis is regulated by a unique bHLH-type transcription factor in Coptis japonica. Plant and Cell Physiology ; doi: 10.1093/pcp/pcr062

Cheong, B-E., Takemura, T., Yoshimatsu, K., Sato, F. (2011) Molecular cloning of an O-methyltransferase from adventitious roots of Carapichea ipecacuanha.  Biosci. Biotechnol. Biochem., 75(1): 107-113.

Koyama T, Mitsuda N, Seki M, Shinozaki K, Ohme-Takagi M.
TCP Transcription Factors Regulate the Activities of ASYMMETRIC LEAVES1 and miR164, as Well as the Auxin Response, during Differentiation of Leaves in Arabidopsis. Plant Cell. 2010 Nov 30. [Epub ahead of print]PMID: 21119060

Morishige, T., Tamakoshi, M., Takemura, T., and Sato, F. (2010) Molecular characterization of O-methyltransferases involved in isoquinoline alkaloid biosynthesis in Coptis japonica. Proc. Jpn. Acad., Ser. B. 86, 757-768. PMID: 20689233 (Free Article)

Yabuta, S., Ifuku, K., Takabayashi, A., Ishihara, S., Ido, K., Ishikawa, N., Endo, T., Sato, F. (2010) Three PsbQ-Like Proteins Are Required for the Function of the Chloroplast NAD(P)H Dehydrogenase Complex in Arabidopsis. Plant and Cell Physiol. 51: 866-876. doi:10.1093/pcp/pcq060. Advance Access published on April 29, 2010.

Takemura T, Ikezawa N, Iwasa K, Sato F. (2010) Metabolic diversification of benzylisoquinoline alkaloid biosynthesis through the introduction of a branch pathway in Eschscholzia californica. Plant and Cell Physiol. 51: 949-959. doi:10.1093/pcp/pcq063. Advance Access published on April 28, 2010.

Ido, K., Ifuku, K., Yamamoto, Y., Ishihara, S., Murakami, A., Takabe, K., Miyake, C., Sato, F. (2009) Knockdown of the PsbP protein does not prevent asembly of the dimeric PSII core complex but impairs accumulation of photosystem II supercomplexs in tobacco. Biochimica Biophysic. Acta. 1787, 873-881.

Ishida S, Takabayashi A, Ishikawa N, Hano Y, Endo T, Sato F. (2009) A novel nuclear-encoded protein, NDH-Dependent cyclic electron Flow 5, is essential for the accumulation of chloroplast NAD(P)H dehydrogenase complexes. Plant Cell Physiol. 2009 Jan ;50(2):383-393.

Ikezawa, N., Iwasa, K. and Sato, F. (2009) CYP719A subfamily of cytochrome P450 oxygenases and isoquinoline alkaloid biosynthesis in Eschscholzia californica. Plant Cell Rep. 28(1), 123-133.

Takabayashi, A., Ishikawa, N., Obayashi, T., Ishida, S., Obokata, J., Endo, T., Sato, F. (2009) Three novel subunits of Arabidopsis chloroplastic NAD(P)H dehydrogenase identified by bioinformatic and reverse genetic approaches. Plant J. 57(2):207-19. Epub 2008 Oct 25.

Minami, H., Kim J-S. Ikezawa, N., Takemura, T., Katayama, T., Kumagai, H., and Sato, F. (2008) Microbial production of plant benzylisoquinoline alkaloids. Proc. Natl. Acad. Sci., USA., 105, 7393-7398.

Ikezawa, N., Iwasa, K., and Sato, F. (2008) Molecular cloning and characterization of CYP80G2, a cytochrome P450 that catalyzes an intramolecular C-C phenol coupling of (S)-reticuline in magnoflorine biosynthesis, from cultured Coptis japonica cells. J. Biol. Chem. 283, 8810-8821 Pubmed/18230623

Ishihara S., Takabayashi A., Ido K., Endo T., Ifuku K., and Sato F., Distinct functions of the two PsbP-like proteins, PPL1 and PPL2, in the chloroplast thylakoid lumen of Arabidopsis thaliana, Plant Physiol., Nov 2007, 145: 668-679. Pubmed/17827269

Minami H., Dubouzet E., Iwasa K., and Sato F., Functional analysis of norcoclaurine synthase in Coptis japonica, J Biol. C hem. 282(9):6274-82 (2007). Epub 2007 Jan 4.

Inui T., Tamura K., Fujii N., Morishige T., and Sato F., Overexpression of Coptis japonica norcoclaurine 6-O-methyltransferase overcomes the rate-limiting step in benzylisoquinoline alkaloid biosynthesis in cultured Eschscholzia californica. Plant Cell Physiol, 48(2):252-62 (2007). Epub 2006 Dec 22. doi: 10.1093/pcp/ PCL062

Ikezawa N, Iwasa, K., and Sato F., Molecular cloning and characterization of methylenedioxy bridge-forming enzymes involved in stylopine biosynthesis in Eschscholzia californica. FEBS J., 274(4):1019-35 (2007)

Kato N., Dubouzet E., Kokabu Y., Yoshida S., Dubouzet J., Yazaki K., and Sato F., Identification of a WRKY protein as a transcriptional regulator of benzylisoquinoline alkaloid biosynthesis in Coptis japonica. Plant Cell Physiol., 48(1):8-18(2007).

Fujii N., Inui T., Iwasa K., Morishige T., and Sato F., Knockdown of berberine bridge enzyme by RNAi accumulates (S)-reticuline and activates a silent pathway in cultured California poppy cells. Transgenic Research, DOI.10.1007/s11248-006-9040-4

Takabayashi, A., Kishine, M., Asada, K., Endo, T., Sato, F., Differential usage of two cyclic electron flows in C4 photosynthesis. Proc. Natl. Acad. Sci.USA., Nov.15, 2005, 102:16898-16903.

Terasaka, K., Blakeslee, J.J., Titapiwatanakun, B., Peer, W.A., Anindita Bandyopadhyay, A., Makam, S.N., Lee, O.R., Richards E.L., , Murphy, A.S., Sato, F., Yazaki, K. (2005), PGP4, an ATP-binding cassette P-glycoprotein, catalyzes auxin transport in Arabidopsis thaliana roots. PLANT CELL 17(11):2922-39 (2005)

Ifuku , K., Yamamoto, Y., Ono, T., Ishihara, S., and Sato, F., PsbP protein, but not PsbQ protein, is essential for the regulation and stabilization of Photosystem II in higher plants, Plant Physiol. 139: 1175-1184.

Ishihara, S., Yamamoto, Y., Ifuku, K., and Sato, F. (2005), Functional analysis of four members of the PsbP family in Photosystem II in Nicotiana tabacum using differential RNA interference. Plant Cell Physiol., 46(12): 1885-1893. 2005

Ohtani, M., Shitan, N., Sakai, K., Martinoia, E., Sato, F., Yazaki, K., Characterization of vacuolar transport of the endogenous alkaloid berberine in Coptis japonica. Plant Physiol., 138: 1939-1946. 2005

Yusuke Kato, Yumiko Yamamoto, Shinya Murakami, Fumihiko Sato, Post-translational regulation of protease activity of CND41 in senescent tobacco leaves. Planta, 222, 643-651(2005)

Joseph Gogo Dubouzet,  Takashi Morishige, Nanae Fujii, Chung-Il An, Ei-ichiro Fukusaki, Kentaro Ifuku, and Fumihiko Sato, Transient RNA Silencing of Scoulerine 9-O-Methyltransferase Expression by Double Stranded RNA in Coptis japonica Protoplasts. Bioscience, Biotechnology, Biochemistry, 69:63-70 (2005)

Endo T, Kawase D, Sato F., Stromal over-reduction by high-light stress as measured by decreases in P700 Oxidation by far-red light and its physiological relevance. Plant Cell Physiol. 46(5):775-81 (2005)

Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F., Functional dissection of two Arabidopsis PsbO proteins, PsbO1 and PsbO2. FEBS J. 272(9): 2165-75 (2005).

Ifuku, K., Nakatsu, T., Shimamoto, R., Yamamoto, Y., Ishihara, S., Kato, H. and Sato, F.,Structure and function of the PsbP protein of photosystem II from higher plants, Photosynthesis Research 2005;84(1-3):251-255 (2005).

Sato, F., RNAi and functional genomics (review), Plant Biotech., 22: 431-442 (2005).

Joseph Gogo Dubouzet, Takashi Morishige, Nanae Fujii, Chung-Il An, Ei-ichiro Fukusaki, Kentaro Ifuku, and Fumihiko Sato, Transient RNA Silencing of Scoulerine 9- O-Methyltransferase Expression by Double Stranded RNA in Coptis japonica Protoplasts. Bioscience, Biotechnology, Biochemistry , 69:63-70 (2005)

Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F., Functional dissection of two Arabidopsis PsbO proteins, PsbO1 and PsbO2. FEBS J . 272(9): 2165-75 (2005) .

Kato, Y., Murakami, S., Yamamoto, Y., Chatani, H., Kondo, Y., Nakano, T.,Yokota, A., and Sato, F. (2004), The DNA-binding protease, CND41, and the degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase in senescent leaves of tobacco. Planta, 220 (1) , 97 - 104, DOI:10.1007/s00425-004-1328-0, PubMed=15252735

Kiminori Takahata, Miyuki Takeuchi, Minoru Fujita, Junichi Azuma, Hiroshi Kamada, Fumihiko Sato (2004), Isolation of putative glycoprotein gene from early somatic embryo of carrot and its possible involvement in somatic embryodevelopment. Plant and Cell Physiology, 45(11):1658-68 (2004), PubMed=15574842

Kishine, M., Takabayashi, A., Munekage, Y., Shikanai, T., Endo, T., and Sato, F., Ribosomal RNA processing and an RNase R family member in chloroplasts of Arabidopsis. Plant Mol. Biol., 55: 595-606 (2004), PubMed=15604703

Ifuku K, Nakatsu T, Kato H, Sato F., Crystal structure of the PsbP protein of photosystem II from Nicotiana tabacum. EMBO Rep. 2004 Apr;5(4):362-7.Epub 2004 Mar 12,PubMed=15031714

Ikezawa N, Tanaka M, Nagayoshi M, Shinkyo R, Sakaki T, Inouye K, Sato F. Molecular cloning and characterization of CYP719, a methylenedioxy bridge-forming enzyme that belongs to a novel P450 family, from cultured Coptis japonica cells. J Biol Chem., 278(40):38557-65, 2003,PubMed=12732624

Koyama, T., Okada, T., Kitajima, S., Ohme-Takagi, M., Shinshi, H., Sato, F. Isolation of tobacco ubiquitin conjugating enzyme cDNA in a yeast two-hybrid system with tobacco ERF3 as bait and its characterization of specific interaction. J.Exp. Bot., 54(385): 1175-1181, 2003,PubMed=12654868

Nakano, T., Kimura, T., Nagata, N., Sekimoto, M., Kawaide, H., Murakami, S., Kaneko, Y., Matsushima, H., Kamiya, Y., Sato, F., and Yoshida, S.: CND41, a chloroplast nucleoid protein, regulated plastid development and dwarfism related to gibberellin biosynthesis. Physiol. Plant. , 117(1):130-136 (2003),PubMed=12610223

Shitan, N., Bazin, I., Dan, K., Obata, K., Kigawa, K., Sato, F., Forestier, C., Yazaki, K., Involvement of CjMDR1, a plant multidrug-resistance-type ATP-binding cassette protein, in alkaloid transport in Coptis japonica. Proc. Natl.Acad.Sci., 100:751-756 (2003),PubMed=_12524452

Ifuku, K., Yamamoto, Y., Sato, F., Specific RNA interference in psbP genes encoded by a multigene family in Nicotinana tabacum with a short 3'-untranslated sequence. Biosci. Biotechnol. Biochem, 67(1); 107-113 (2003),PubMed=12619681

Ifuku, K., and Sato, F., A truncated mutant of the extrinsic 23-kDa protein that absolutely requires the extrinsic 17-kDa protein to function in photosystem II" Plant Cell Physiol., 43 (10):1244-1249 (2002), PubMed=12407205

Koizumi, N., Toyota, K., Kitajima, S., Yamada, Y., Sato, F.; Genomic structure and promoter analysis of phosphoenolpyruvate carboxylase in a C3 plant, Nicotiana sylvestris. Biosci. Biotechnol. Biochem, 66:1691-1696 (2002), PubMed=12353629

Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F., Characterization of an Arabidopsis thaliana mutant with impaired psbO, one of two genes encoding extrinsic 33-kDa proteins in photosystem II. FEBS Letters, 523: 138-142 (2002), PubMed=12123820

Choi KB, Morishige T, Shitan N, Yazaki K, and Sato F.: Molecular cloning and characterization of coclaurine N-methyltransferase from cultured cells of Coptis japonica. J Biol Chem., 277:830-835, 2002, PubMed=11682473

Ifuku K, Sato F.: Importance of the N-terminal sequence of the extrinsic 23 kDa polypeptide in Photosystem II in ion retention in oxygen evolution. Biochim Biophys Acta 2001 Mar 9;1546(1):196-204 PubMed=11257522

Sato, F., Hashimoto, T., Hachiya, A., Tamura, K., Choi, K.-B., Morishige, T., Fujimoto, H. and Yamada, Y.: Metabolic engineering of plant alkaloid biosynthesis. Proc. Natl. Acad. Sci. USA 98(1); 367-372, 2001 PubMed=11124522

Fukushima, E., Arata, Y., Endo, T., Sonnewald, U. and Sato, F.: Improved salt tolerance of transgenic tobacco expressing apoplastic yeast-derived invertase. Plant Cell Physiol. 42(2); 245-249, 2001 PubMed=11230581

Murakami, S., Kondo, Y., Nakano,T. & Sato, F.: Protease activity of CND41, a chloroplast nucleoid DNA-binding protein, isolated from cultured tobacco cells. FEBS Lett. 468;15-18, 2000

Kitajima, S., Koyama, T., Ohme-Takagi, M., Shinshi, H. and Sato, F.: Characterization of gene expression of ERFs, transcription factors of basic PR genes from Nicotiana sylvestris. Plant Cell Physiol. 41 (1); 817-824, 2000

Morishige, T., Tsujita, T., Yamada, Y. and Sato, F.: Molecular characterization of the S-adenosyl-L-methionine: 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase in isoquinoline alkaloid biosynthesis in Coptis japonica. J. Biol. Chem. 275; 23398-23405, 2000

Books and reviews

Year 2010

Mizutani M, Sato F. (2010) Unusual P450 reactions in plant secondary metabolism. Arch Biochem Biophys. 2010 Oct 23. [Epub ahead of print] PMID: 20920462 [Review]

Minami H, Ikezawa N, Sato F. (2010)
Microbial expression of alkaloid biosynthetic enzymes for characterization of their properties. Methods Mol Biol. 643:111-120. PMID: 20552447

Takemura T, Chow YL, Todokoro T, Okamoto T, Sato F. (2010)
Over-expression of rate-limiting enzymes to improve alkaloid productivity. Methods Mol Biol. 643:95-109. PMID: 20552446

Yamada Y, Kato N, Kokabu Y, Luo Q, Dubouzet JG, Sato F. (2010)
Identification of regulatory protein genes involved in alkaloid biosynthesis using a transient RNAi system. Methods Mol Biol. 643:33-45. PMID: 20552442

Year2007

Sato Fumihiko, Inui Takayuki, and Takemura Tomoya (2007) Metabolic Engineering in Isoquinoline Alkaloid Biosynthesis. Current Pharmaceutical Biotechnology, Aug;8(4):211-8.

Sato Fumihiko, Inai Koji, and Hashimoto Takashi (2007)  Metabolic Engineering in Alkaloid Biosynthesis: Case Studies in Tyrosine- and Putrescine-Derived Alkaloids. In: Applications of Plant Metabolic Engineering, Chapter 6 (Editors: Prof. Verpoorte Robert, Prof. Alfermann A. W. and Dr. Johnson T. S.), Springer, ISBN: 978-1-4020-6030-4, pp.145-173

Year 2003

Sato, F., Secondary metabolites and biotechnology in plants, In "Agrobiotechnology and Plant Tissue Culture", ed. S.S.Bhojwani and W.-Y. Soh, Science Pub. Inc., Enfield (NH), USA, ISBN 1-57808-243-9, pp97-104 (2003)

Year 2000

Sato, F.: Chloroplast function and plant development, Tanpakushitsu-Kakusan-Koso, 45; 147-152, 2000 (in Japanese)

Sato, F.: Introduction to molecular and cellular biology of chloroplast biogenesis, Tanpakushitsu-Kakusan-Koso, 45;105-1107, 2000 (in Japanese)

Sato, F., Kitajima, S., Koyama, T., Okada, T., Ohme-Takagi, M. and Shishi, H.: Regulation of PR-5d (osmotin-like protein) gene expression via ERFs (ethylene responsive factors). In The 14th Symposium on Plant Biotechnology: 2000 Japan-Korea Joint Symposium, Vol. 14: 103-109 , 2000.

Sato, F.:Molecular and cellular biology of plant functions using photoautotrophic cultured cells and its application, Chemical regulation of plants, 35(1);8-16, 2000 (in Japanese)

Sato, F.: GM foods, Medical Practice, 17(10); 1788-1790, 2000 (in Japanese)

Year 1999

Kitajima, S. & Sato, F.: Plant pathogeniesis-related proteins: molecular mechanisms of gene expression and protein function. J.Biochem., 125(1): 1-8 (1999)

B. Educational Activities

B-1. On-campus teaching

a) Courses given

Undergraduate level: Outline of Cell Physiology (Sato), Laboratory Course in Plant Biochemistry (Endo, Ifuku), Introduction to Research (Sato, Endo)

Graduate level: Experimental Course of Molecular and Cellular Biology (Sato, Endo), Molecular and Cellular Biology Seminar (Sato, Endo)