Studies of phytochromes and signal transductbn pathways in cyanobacterial phototaxis

• Project/Area Number: 13440235
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 植物生理
• Research Institution: The University of Tokyo
• Principal Investigator: IKEUCHI Masahiko The University of Tokyo, Graduate School of Arts and Sciences, Professor, 大学院・総合文化研究科, 教授 (20159601)
• Project Period (FY): 2001 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥14,700,000 (Direct Cost: ¥14,700,000); Fiscal Year 2003: ¥4,600,000 (Direct Cost: ¥4,600,000); Fiscal Year 2002: ¥4,600,000 (Direct Cost: ¥4,600,000); Fiscal Year 2001: ¥5,500,000 (Direct Cost: ¥5,500,000)
• Keywords: cyanobacteria / blue-green algae / phototaxis / flavin sensor / photoconversion / phytochrome / photoreceptor / phycocyanin / 運動性 / 形質転換能 / 分割遺伝子

Research Abstract

A gene for a novel flavin-binding protein that is involved in positive phototaxis was cloned from T. elongatus and expressed in E. coli, and expressed. The protein was crystallized and resolved at 2.0 A.
Novel phytochrome-like gene pixA was identified as a putative photoreceptor for positive phototaxis. Regulator proteins encoded by nixB and nixC are hypothesized by combinatorial mutagenesis.
Regulator genes essential for assembly of motility pili were identified as follows : pilG, pilH, pill, pilJ, pilL-N and pilL-C. The latter two genes were separately expressed on distinct loci on the chromosome.
Positive phototaxis photoreceptor PixJ1 protein was isolated from Synechocystis sp. PCC 6803. The protein thus isolated showed novel absorption spectra and difference spectrum in blue and green light regions. The chromophore of PixJ1 was concluded to be a derivative of phycocyanobilin.
Four novel flavin-binding photoreceptor proteins were expressed heterologously and their photocycles were partially characterized.
The PAS domain proteins of a filamentous cyanobacterium Anabaena sp. PCC 7120 were systematically studied by bioinformatics and biochemical analyses.
Genes for the pili subunits were systematically disrupted and identified as pil9,pi110, and pil11.

Research Products

• [Journal Article] Improved genetic transformation of the thermophilic cyanobacterium, Thermosynechococcus elongatus BP-1 (2004)
  • Author(s): Iwai et al.
  • Journal Title: Plant Cell Physiology 45 Pages: 171-175
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Response to oxidative stress involves a novel peroxiredoxin gene in the unicellular cyanobacterium Synechocystis sp. PCC 6803 (2004)
  • Author(s): Kobayashi et al.
  • Journal Title: Plant Cell Physiology 45 Pages: 290-299
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Use of segment-based microarray in the analysis of global gene expression in response to various environmental stresses in the cyanobacterium Anabaena sp. PCC 7120 (2004)
  • Author(s): Sato et al.
  • Journal Title: J. Gen. Appl. Microbiol. 50 Pages: 1-8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Molecular evolution of PAS domain-containing proteins of filamentous cyanobacteria through the domain shuffling and domain duplication (2004)
  • Author(s): Narikawa et al.
  • Journal Title: DNA Research 11 Pages: 69-81
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Iwai, M., Katoh, H., Katayama, M. and Ikeuchi, M. Improved genetic transformation of the thermophilic cyanobacterium, Thermosynechococcus elongatus BP-1. (2004)
  • Author(s): Iwai, M., Katoh, H., Katayama, M., Ikeuchi, M.
  • Journal Title: Plant Cell Physiol 45 Pages: 171-175
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Response to oxidative stress involves a novel peroxiredoxin gene in the unicellular cyanobacterium Synechocystis sp. PCC 6803. (2004)
  • Author(s): Kobayashi, M., Ishizuka, T., Katayama, M., Kanehisa, M., Bhattacharyya-Pakrasi, M., Pakrasi, H.B., Ikeuchi, M.
  • Journal Title: Plant Cell Physiol. 45 Pages: 290-299
  • NAID: 10012724002
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Use of segment-based microarray in the analysis of global gene expression in response to various environmental stresses in the cyanobacterium Anabaena sp. PCC 7120. (2004)
  • Author(s): Sato, N., Ohmori, M., Ikeuchi, M., Tashiro, K., Wolk, P.C., Kaneko, T., Okada, K., Tsuzuki, M., Ehira, S., Katoh, H., Okamoto, S., Yoshimura, H., Fujisawa, T., Kamei, A., Yoshihara, S., Narikawa, R., Hamano, T., Tabata, S., Kuhara, S.
  • Journal Title: J.Gen.Appl.Microbiol. 50 Pages: 1-8
  • NAID: 130004449126
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular evolution of PAS domain-containing proteins of filamentous cyanobacteria through the domain shuffling and domain duplication. (2004)
  • Author(s): Narikawa, R., Okamoto, S., Ikeuchi, M., Ohmori, M.
  • Journal Title: DNA Res. 11 Pages: 69-81
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stable Positional Cloning of Long Continuous DNA in the Baclllus subti is genome vector (2003)
  • Author(s): Itaya et al.
  • Journal Title: J. Biochemistry 134 Pages: 513-519
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Biochemical and functional characterization of a eukaryotic-type protein kinase, SpkB, in the eyanobacterium, Synechocystis sp. PCC 6803 (2003)
  • Author(s): Kamei et al.
  • Journal Title: Curr Microbiol 46 Pages: 296-301
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stable positional cloning of long continuous DNA in the Bacillus subtilis genome vector. (2003)
  • Author(s): Itaya, M., Fujita, K., Ikeuchi, M., Koizumi, M., Tsuge, K.
  • Journal Title: J.Biochem. 134 Pages: 513-519
  • NAID: 10012058225
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Biochemical and functional characterization of a eukaryotic-type protein kinase, SpkB, in the cyanobacterium, Synechocystis sp. (2003)
  • Author(s): Kamei, A., Yoshihara, S., Yuasa, T., Geng, X., Ikeuchi, M.
  • Journal Title: PCC 6803.Curr.Microbiol. 46 Pages: 296-301
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Post-genome project of cyanobacteria. (2001)
  • Author(s): Ohmori, M., M.Ikeuchi
  • Journal Title: Tanpakushitu Kakusann Kouso 46 Pages: 2486-2492
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] タンパク質・核酸・酵素(分担) (2001)
  • Author(s): 大森正之・池内昌彦
  • Total Pages: 2492
  • Publisher: シアノバクテリアの光合成遺伝子・タンパク質システムの解明
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Itaya et al.: "Stable Positional Cloning of Long Continuous DNA in the Bacillus subtilis genome vector"J.Biochemistry. 134. 513-519 (2003)
• [Publications] Narikawa et al.: "Newly identified motifs within PAS domains of filamentous cyanobacteria"Genome Informatics. 14. 444-445 (2003)
• [Publications] Iwai et al.: "Improved genetic transformation of the thermophilic cyanobacterium, Therm osynechococcus elongatus BP-1"Plant Cell Physiology. 45. 171-175 (2004)
• [Publications] Kobayashi et al.: "Response to oxidative stress involves a novel peroxiredoxin gene in the unicellular cyanobacterium Synechocystis sp. PCC 6803"Plant Cell Physiology. 45(In press). (2004)
• [Publications] Sato et al.: "Use of segment-based microarray in the analysis of global gene expression in response to various environmental stresses in the cyanobacterium Anabaena sp.PCC 7120"J.Gen.Appl.Microbiol.. (In press). (2004)
• [Publications] Narikawa et al.: "Molecular evolution of PAS domain-containing proteins of filamentous cyanobacteria through the domain shuffling and domain duplication"DNA Research. (In press). (2004)
• [Publications] Yoshimura, H. et al.: "A cAMP receptor protein, SYCRP1, is responsible for the cell motility of Synechocystis sp. PCC 6803"Plant Cell Physiology. 43. 460-463 (2002)
• [Publications] Yoshihara, S. et al.: "pilG gene cluster and split pilL genes involved in pilus biogenesis, motility and genetic transformation in the cyanobacterium Synechocystis sp. PCC 6803"Plant Cell Physiology. 43. 513-521 (2002)
• [Publications] Kamei, A. et al.: "Biochemical examination of the potential eukaryotic-type protein kinase genes in the complete genome of the unicellular cyanobacterium Synechocystis sp. PCC 6803"DNA Research. 9. 71-78 (2002)
• [Publications] Nakamura, Y. et al.: "Complete genome structure of the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1"DNA Research. 9. 123-130 (2002)
• [Publications] Kashino, Y. et al.: "Low-molecular-mass polypeptide components of a photosystem II preparation from the thermophilic cyanobacterium Thermosynechococcus vulcanus"Plant Cell Physiology. 43. 1366-1373 (2002)
• [Publications] Kamei, A. et al.: "Biochemical and functional characterization of a eukaryotic-type protein kinase, SpkB, in the cyanobacterium, Synechocystis sp. PCC 6803"Current Microbiology. (in press). (2003)
• [Publications] Hihara, Y. et al.: "DNA microarray analysis of cyanobacterial gene expression during acclimation to high light"Plant Cell. 13. 793-806 (2001)
• [Publications] Katoh, H. et al.: "Functional analysis of psbV and a novel c-type cytochrome gene psbV2 of the thermophilic cyanobacterium Thermosynechococcus elongatus strain BP-1"Plant and Cell Physiology. 42. 599-607 (2001)
• [Publications] Ikeuchi, M., Tabata, S.: "Synechocystis sp. PCC 6803 - a useful tool in the study of the genetics of cyanobacteria"Photosynthesis Research. 70. 73-83 (2001)
• [Publications] Ohmori, M. et al.: "Characterization of genes encoding multi-domain proteins in the genome of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120"DNA Research. 8. 271-284 (2001)
• [Publications] Yoshimura, H. et al.: "A cAMP receptor protein, SYCRP1, is responsible for the cell motility of Synechocystis sp. PCC 6803"Plant and Cell Physiology. (印刷中).
• [Publications] Yoshihara, S. et al.: "pilG gene cluster and split pilL genes involved in pilus biogenesis, motility and genetic transformation in the cyanobacterium Synechocystis sp. PCC 6803"Plant and Cell Physiology. (印刷中).
• [Publications] 大森正之・池内昌彦: "タンパク質・核酸・酵素(分担)(シアノバクテリアの光合成遺伝子・タンパク質システムの解明)"2492 (2001)