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SecY functions that support the dynamic movement of SecA, a protein-translocating ATPase

Research Project

Project/Area Number 09480150
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

ITO Koreaki  Kyoto Univ., Inst. Virus Res., Professor, ウイルス研究所, 教授 (90027334)

Co-Investigator(Kenkyū-buntansha) MORI Hiroyuki  Kyoto Univ., Inst. Virus Res., Instructor, ウイルス研究所, 助手 (10243271)
AKIYAMA Yoshinori  Kyoto Univ., Inst. Virus Res., Associate Professor, ウイルス研究所, 助教授 (10192460)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 1999: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 1998: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
Keywordsprotein translocation / SecY / SecA / membrane protein / protein-protein interaction / ATPase
Research Abstract

We studied protein interactions in the cellular machinery that mediates protein translocation across the E. coli cytoplasmic membrane. We found that in the membrane-embedded SecY-SecE-SecG complex of protein translocase, each of SecE and SecG interacts independently with SecY, the central submit. The residue 240 of SecY is particularly important for the SecY's interaction with SecE. We established a system, using a SecY-interaction protein, Syd, and a secY mutation that impairs SecY-SecE interaction, in which we can manipulate the SecA-binding ability of the SecYEG channel complex. SecA, the preprotein-driving ATPase, inserts into the membrane in response to ATP and a preprotein and deinserts from the membrane upon hydrolysis of ATP, thereby facilitating the preprotein movement into the membrane, We identified a secY mutation(SecY205)that impairs the SecA insertion reaction, and suggested that it alters the translocation channel such that the altered channel cannot accept a SecA molecu … More le that is in complex with the preprotein. We isolated suppressor mutations in SecA that overcame the SecY205 defect, and characterized the mutant proteins biochemically. These studies suggested that SecY interacts specifically with SecA to allow the insertion of preprotein-SecA complex, and the SecA insertion reactions is indeed important for protein translocation both in vivo and in vitro. Our mutant analyses suggest that the regions around the two Walker A motifs in SecA are important for the functional SecA interaction, while the secondary ATP-binding region is important for the regulation of the SecA functions. Some SecA mutations suppressed the SecG deletion mutation and our analyses suggest that SecG has a role of assisting SecA and its effective utilization of ATP for protein translocation. We have shown that SecA is activated by its interaction with SecY, and that the fifth cytoplasmic domain of SecY is important for the SecA activation. We identhified particular residues in this domain of SecY, which is crucial for the SecA activation. Less

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (57 results)

All Other

All Publications (57 results)

  • [Publications] Ito,K., Matsuo,E.. And Akiyama,Y.: "A class of integral membrane proteins will be overlooked by the "proteome" study that is based on two-dimensional gel electrophoresis"Mol.Microbiol.. 31. 1600-1601 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama,Y.: "Self-processing of FtsH and its implication for the cleavage specificity of this protease"Biochemistry. 38. 11693-11699 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kobayashi,T. and Ito,K.: "Respiratory chain strongly oxidizes the CXXC motif of DsbB in the Escherichia coli disulfide-bond formation pathway"EMBO J.. 18. 1192-1198 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara,A., Akiyama,Y. and Ito,K.: "Dislocation of membrane proteins in FtsH-mediated proteolysis"EMBO J.. 18. 2970-2981 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo,E., Sampei,G., Mizobuchi,K. and Ito,K.: "The plasmid F OmpP protease, a homologue of OmpT, as a potential obstacle to E. coli-based protein production"FEBS Lett.. 461. 68 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo,E.. and Ito,K.: "Genetic analysis of an essential cytoplasmic domain of Escherichia coli Sec Y based on resistance to Syd, a Sec Y-interacting protein"Mol.Gen.Genet.. 258. 240-249 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo,E.., Mori,H., Shimoike,T. and Ito,K.: "Syd, a Sec Y-interacting protein, excludes Sec A from the Sec YE complex with an altered Sec Y24 subunit"J.Biol.Chem.. 273. 18835-18840 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsumoto,G.., Mori, and Ito,K.: "Role of Sec G in ATP-and Sec A-dependent protein translocation"Proc.Natl.Acad.Sci.USA. 95. 13567-3572 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara,A. and Ito,K.: "Translocation, folding and stability of the HflKC complex with signal-anchor topogenic sequences"J.Biol.Chem.. 273. 29770-29775 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara,A., Akiyama,Y. and Ito,K.: "Different pathways for protein degradation by the FtsH/HflKC membrane-embedded protease complex : an implication from the interference by a mutant form of a new substrate protein, YccA"J.Mol.Biol.. 279. 175-188 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama,Y., Ehrmann,M., Kihara,A. and Ito,K.: "Polypeptide-binding of E. coli FtsH"Mol.Microbiol.. 28. 803-812 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama,Y., Kihara,A., Mori,H., Ogura,T. and Ito,K.: "Roles of periplasmic domain of Escherichia coli FtsH (HflB) in protein interactions and activity modulation"J.Biol.Chem.. 273. 22326-22333 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Homma,T., Yoshihisa,T. and Ito,K.: "Subunit interactions in the Escherichia coli protein translocase:SecE and SecG associate independently with SecY"FEBS Lett.. 408. 11-15 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Taura,T.,Yoshihisa,T. and Ito,K: "Protein translocation functions of Escherichia coli Sec Y : in vitro characterization of cold-sensitive sec Y mutants"Biochimie. 79. 517-521 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsumoto,G.., Yoshihisa,T. and Ito,K.: "Sec Y and Sec A interact to allow Sec A insertion and protein translocation across the Escherichia. Coli plasma membrane"EMBO J.. 16. 6384-6393 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara,A., Akiyama,Y. and Ito,K.: "Host regulation of lysogenic decision bacteriohage λ : transmembrane modulation of FtsH(HflB), the cII degrading protease, by HflKC(HflA)"Proc.Natl.Acad.Sci.USA. 94. 5544-5549 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sone,M., Kishigami,S., Yoshihisa,T. and Ito,K.: "Roles of bonds in bacterial alkaline phosphatase"J.Biol.Chem.. 272. 6174-6178 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sone,M., Akiyama,Y. and Ito,K.: "Differential in vivo roles played by DsbA and DsbC in the formation of protein disulfide bonds"J.Biol.Chem.. 272. 10349-10352 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kobayashi,T., Kishigami,S., Sone,M., Inokuchi,H., Mogi,T. and Ito,K.: "Respiratory electron transfer chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells"Proc.Natl.Acad.Sci.USA. 94. 11857-11862 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Homma, T., Yoshihisa, T. and Ito, K: "Subunit interactions in the Escherichia coli protein translocase : SecE and SecG associate independently with SecY."FEBS Lett. 408. 11-15 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Taura, T., Yoshihisa, T. and Ito, K: "Protein translocation functions of Escherichia coli SecY : in vitro characterization of cold-sensitive secY mutants"Biochimie. 79. 517-521 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsumoto, G., Yoshihisa, T. and Ito, K: "SecY and SecA interact to allow SecA insertion and protein translocation across the Escherichia. coli plasma membrane."EMBO J.. 16. 6384-6393 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara, A., Akiyama, Y. and Ito, K.: "Host regulation of lysogenic decision in bacteriohage λ: transmembrane modulation of FtsH (HflB), the cII degrading protease, by HflKC (HflA)"Proc. Natl. Acad. Sci USA. 94. 5544-5549

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sone, M., Kishigami, S., Yoshihisa, T. and Ito, K: "Roles of disulfide bonds in bacterial alkaline phosphatase."J. Biol. Chem. 272. 6174-6178 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sone, M., Akiyama, Y.and Ito, K: "Differential in vivo roles played by DsbA and DsbC in the formation of protein disulfide bonds."J. Biol. Chem. 272. 10349-10352 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kobayashi, T., Kishigami, S., Sone, M., Inokuchi, H., Mogi, T. and Ito, K: "Respiratory electron transfer chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells"Proc. Natl. Acad. Sci. USA. 94. 11857-11862 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara, A., Akiyama, Y.and Ito, K: "Different pathways for protein degradation by the FtsH/HflKC membrane-embedded protease complex : an implication from the interference by a mutant form of a new substrate protein, YccA."J. Mol. Biol. 279. 175-188 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama, Y., Ehrmann, M., Kihara, A. and Ito, K.: "Polypeptide-binding of E. coli FtsH."Mol. Microbiol. 28. 803-812 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo, E. and Ito, K: "Genetic analysis of an essential cytoplasmic domain of Escherichia coli SecY based on resistance to Syd, a SecY-interacting protein."Mol. Gen. Genet.. 258. 240-249 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo, E., Mori, H., Shimoike, T. and Ito, K: "Syd, a SecY-interacting protein, excludes SecA from the SecYE complex with an altered SecY24 subunit."J. Biol. Chem. 273. 18835-18840 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama, Y., Kihara, A., Mori, H., Ogura, T. and Ito, K: "Roles of the periplasmic domain of Escherichia coli FtsH (HflB) in protein interactions and activity modulation."J. Biol. Chem. 273. 22326-22333 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara, A. and Ito, K: "Translocation, folding and stability of the HflKC complex with signal-anchor topogenic sequences."J. Biol. Chem. 273. 29770-29775

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsumoto, G., Mori, H. and Ito, K: "Roles of SecG in ATP-and SecA-dependent protein translocation."Proc. Natl. Acad. Sci. USA. 95. 13567-13572 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Ito, K., Matsuo, E. and Akiyama, Y: "A class of integral membrane proteins will be overlooked by the "proteome" study that is based on two-dimensional gel electrophoresis"Mol. Microbiol. 31. 1600-1601 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akiyama, Y.: "Self-processing of FtsH and its implication for the cleavage specificity of this protesase."Biochemistry. 38. 11693-11699 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kobayashi, T. and Ito, K: "Respiratory chain strongly oxidizes the CXXC motif of DsbB in the Escherichia coli disulfide-bond formation pathway."EMBO J.. 18. 1192-1198 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kihara, A. Akiyama, Y. and Ito, K: "Dislocation of membrane proteins in FtsH-mediated proteolysis"EMBO J.. 18. 2970-2981 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo, E., Sampei, G., Mizobuchi, K. and Ito, K: "The plasmid F OmpP protease, a homologue of OmpT, as a potential obstacle to E. coli-based proten production"FEBS Lett. 461. 6-8 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Ito K.,Matsuo E.and Akiyama Y.: "A class of integral membrane proteins will be overlooked by the "proteome" study that is based on two-dimensional gel electrophoresis"Mol. Microbiol.. 31. 1600-1601 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Akiyama Y.: "Self-processing of FtsH and its implication for the cleavage specificity of this protease"Biochemistry. 38. 11693 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kobayashi T.and Ito K.: "Respiratory chain strongly oxidizes the CXXC motif of DsbB in the Escherichia coli disulfide-bond formation pathway"EMBO J.. 18. 1192-1198 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kihara A.,Akiyama Y.and Ito K.: "Dislocation of membrane proteins in FtsH-mediated proteolysis"EMBO J.. 18. 2970-2981 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Matsuo E.,Sampei G.,Mizobuchi K.and Ito K.: "The plasmid F OmpP protease, a homologue of OmpT, as a potential obstacle to E.Coli-based protein production"FEBS Lett.. 461. 6-8 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Matsuo,E.and Ito,K.: "Genetic analysis of an essential cytoplasmic domain of Escherichia coli SecY based on resistance to Syd,a SecY-interacting protein." Mol.Gen.Genet.258. 240-249 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Matsuo,E.Mori,H.,Shimoika,T.and Ito,K.: "Syd,a SecY-interacting protein,excludes SecA from the SecYE complex with an altered SecY24 subunit." J.Biol.Chem.273. 18835-18840 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Matsumoto,G.,Mori,H.and Ito,K.: "Roles of SecG in ATP-and SecA-dependent protein translocation." Proc.Natl.Acad.Sci.USA. 95. 13567-13572 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kihara,A.and Ito,K.: "Translocation,folding and stability of the HflKC complex with signal-anchor topogenic sequences." J.Biol.Chem.273. 29770-29775 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kihara,A.,Akiyama,Y.and Ito,K.: "Different pathways for protein degradation by the EtsH/HflKC membrane-embedded protease complex:an implication from the interference by a mutant form of a new substrate protein,YccA." J.Mol.Biol.279. 175-188 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Akiyama,Y.,Ehrmann,M.,Kihara,A.and Ito,K.: "Polypeptide-binding of E.coli FtsH." Mol.Microbiol.28. 803-812 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Akiyama,Y.,Kihara,A.,Mori,H.,Ogura,T.and Ito,K.: "Roles of the periplasmic domain of Escherichia coli FtsH(HflB) in protein interactions and activity modulation." J.Biol.Chem.273. 22326-22333 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Homma,T., Yoshihisa,T. and Ito,K.: "Subunit interactions in the Escherichia coli protein translocase : SecE and SecG associate independently with SecY." FEBS Lett.408. 11-15 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Taura,T., Yoshihisa,T. and Ito,K.: "Protein translocation functions of Escherichia coli SecY : in vitro characterization of cold-sensitive secY mutants." Biochimie. 79. 517-521 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Matsumoto,G., Yoshihisa,T. and Ito,K.: "SecY and SecA interact to allow SecA insertion and protein translocation across the Escherichia. Coli plasma membrane." EMBO J.16. 6384-6393 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kihara,A., Akiyama,Y. and Ito,K.: "Host regulation of lysogenic decision in bacteriohage λ : transmembrane modulation of FtsH (HflB),the cII degrading protease,by HflKC (HflA)." Proc.Natl.Acad.Sci USA. 94. 5544-5549 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sone,M., Kishigami,S., Yoshihisa,T. and Ito,K.: "Roles of disulfide bonds in bacterial alkaline phosphatase." J.Biol.Chem.272. 6174-6178 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sone,M., Akiyama,Y. and Ito,K.: "Differential in vivo roles played by DsbA and DsbC in the formation of protein disulfide bonds." J.Biol.Chem.272. 10349-10352 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi,T., Kishigami,S., Sone,M., Inokuchi,H., Mogi,T. and Ito,K.: "Respiratory electron transfer chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically grwoing Escherichia coli cells." Proc.Natl.Acad.Sci. USA. 94. 11857-11862 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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