Protein translocation across the cytoplasmic membrane of bacteria

1996 Fiscal Year Final Research Report Summary

• Project/Area Number: 07308069
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 総合
• Research Field: Structural biochemistry
• Research Institution: University of Tokyo
• Principal Investigator: TOKUDA Hajime University of Tokyo, Institute of Molecular and Cellular Biosciences, Professor, 分子細胞生物学研究所, 教授 (40125943)
• Co-Investigator(Kenkyū-buntansha): WATANABE Makoto Oita Medical University, Department of Biochemistry, Professor, 医学部, 教授 (20142408) ; YAMANE Kunio University of Tsukuba, Institute of Biological Sciences, Professor, 生物科学系, 教授 (20013336) ; HIRATA Hajime Institute of Technology, Faculty of Science, Professor, 理学部, 教授 (40049052) ; ENDO Toshiya Nagoya University, Faculty of Science, Professor, 理学部, 教授 (70152014) ; ITO Koreaki Kyoto University, Institute of Virus Research, Professor, ウィルス研究所, 教授 (90027334)
• Project Period (FY): 1995 – 1996
• Keywords: Protein translocation / Prokaryote / Reconstitution / SecY / SecE / SecG / SecA

Research Abstract

1. SecG was found to interact with SecA and facilitate the membrane insertion-deinsertion cycle of SecA.
2. Mode of interaction of SecY-SecE-SecG complex was revealed.
3. SecA was found to be involved in the import of cytochrome f into thylakoid.
4. SecY and SecE were reconstituted into a planar lipid bilayr membrane and found to form a ion-conducting channel.
5. Step-wise translocation of precursor protein was found to be caused by a hydrophobic region of the precursor.
6. Structure and function of Bacillus subtilis SRP homologue were revealed.
7. Highly active protein translocation system was reconstituted into proteoliposomes.

Research Products

• [Publications] Nishiyama,K.: "Inversion of the membranc topology of SecG coupled with SecA-dependent preprotein translocation." Cell. 85. 71-81 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kontinen,V.P: "Roles of the conserved cytoplasmic region and non-conserved carboxyterminal region of SecE in E. coli protein translocace." J.Biochemistry. 119. 1124-1130 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kontinen,V.P.: "The secG deletion mutation of Escherichia coli is suppressed by expression of a novel regulatory gene of Bacillus subtilis." FEBS Lett.389. 281-284 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hirano,M: "The carboxyl-terminal region is essential for SecA dimerization." Biochem.Biophys.Res.Commun.229. 90-95 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Akiyama,Y.: "FtsH (HflB) is an ATP-dependent protease selectively acting on SecY and some other membrane proteins." J.Biol.Chem.271. 31196-31201 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Homma,T.: "Intracellular stability of a fragment of β-galactosidase : effects of aminoterminally fused polypeptides." Biochem.Biophys.Res.Commun.215. 452-458 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Akiyama,Y.: "Subunit a of proton ATPase F_0 sector is a substrate of the FtsH protease in Escherichia coli." FEBS Lett.399. 26-28 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kishigami,S.: "Roles of cysteine residues of DsbB in its activity to reoxidize DsbA,the protein disulfide bond catalyst of Escherichia coli." Genes to Cells. 1. 201-208 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sone,M.: "Roles of disulfide bonds in bacterial alkaline phosphatase." J.Biol.Chem.(in press). (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Endo,T.: "Mitochondrial presequences can induce aggregation of unfolded proteins." FEBS Lett.359. 93-96 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nohara,T.: "Isolation and charac terization of the cDNA for pea chloroplast SecA : evolutionary conservation of the bacterial-type SecA-dependent protein transport within chloroplasts." FEBS Lett.364. 305-308 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Endo,T.: "Avidin fusion protein as a tool to generate a stable translocation intermediate spanning the mitochondrial membranes." J.Biochem.118. 753-759 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Endo,T.: "Binding of mithochondrial presequences to yeast cytosolic heat shock protein 70 depends of the amphiphilicity of the presequence." J.Biol.Chem.271. 4161-4167 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kanamori,T.: "Probing the environment along the protein import pathways in yeast mitochondria by site-specific photocrosslinking." Proc.Natl.Acad.Sci.U.S.A.94. (in press). (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Murai,N.: "A Novel Insertion Sequence (SI)-like Element of the Thermophilic Bacterium PS3 promotes Expression of the Alanine Carrier Protein-encoding Gene." Gene. 163. 103-107 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Jin,T.: "Selective Na+Transport through Phospholipid Bilayer Membrane by a Synthetic Calix〔4〕arene Carrier." Langmuir. 12. 2684-2689 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kamata,H.: "Nerve Growth Factor and Forskolin prevent H_2O_2-induced Apoptosis in PC12 Cells by Glutathione Independent Mechanism." Neurosci.Lett.212. 179-182 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kamata,H.: "Involvement of Cellular Redox State in Cell Survival and Neuronal Differentiation in PC12 Cells." Biochem.Soc.Trans.24. 541 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kamata,H.: "Suppression of NGF-induced Neuronal Differentiation of PC12 Cells ; N Acetylcysteine (NAC) uncouples the Signal Transduction from Ras to the MAP Kinase Cascade." J.Biol.Chem.271. 33018-33025 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Uchida,K.: "Stepwise Movement of Preproteins in the Process of Translocation across the Cytoplasmic Membrance of Escherichia coli." J.Biol.Chem.270. 30862-30868 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sato,K.: "Characterization of a Potential Catalytic Residue,Asp-133,in the High Affinity ATP-binding Site of Escherichia coli SecA,Translocation ATPase." J.Biol.Chem.271. 17439-17444 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Sato,K.: "Short hydrophobic segments in the mature domain of proOmpA determine its stepwise movement during translocation across the cytoplasmic membrane of Escherichia coli" J.Biol.Chem.272(in press). (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nakane,A.: "Acquisition of azide-resistance by clevated SecA ATPase activity confers azide-resistance upon cell growth and protein translocation in Bacillus subtilis." Microbiology. 141. 113-121 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ogawa,K.: "The nasB operon and nasA gene are required for nitrate/nitrite assimilation in Bacillus subtilis." J.Bacteriol.177. 1409-1413 (1995)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kurita,K.: "Identification of a region of Bacillus subtilis Ffh,a homologue of mammalian SRP54 protein,that is essential for binding to small cytoplasmic RNA." J.Biol.Chem.271. 13140-13146 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Yamane,K.: "25^0-36^0 region of the Bacillus subtilis chromomose : determination of the sequence of a 146 kb segment and identification of 113 genes." Microbiology. 142. 3047-3056 (1996)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Watanabe,M.: "High-affinity binding of Escherichia coli SecB to the signal sequence region of a presecretory protein" Proc,Natl.Acad.Sci.USA. 92. 10133-10136
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nishiyama, K., Suzuki, T., and Tokuda, H.: "Inversion of the membrane topology of SecG coupled with SecA-dependent preprotein translocation." Cell. 85. 71-81 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kontinen, V.P., Yamanaka, M., Nishiyama, K., and Tokuda, H.: "Roles of the conserved cytoplasmic region and non-conserved carboxy-terminal region of SecE in E.coli protein translocase." J.Biochemistry. 119. 1124-1130 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kontinen, V.P., Helander, I.M., and Tokuda, H.: "The secG deletion mutation of Escherichia coli is suppressed by expression of a novel regulatory gene of Bacillus subtilis." FEBS Lett.389. 281-284 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hirano, M., Matsuyama, S., and Tokuda, H.: "The carboxyl-terminal region is essential for SecA dimerization." Biochem. Biophys. Res. Commun.229. 90-95 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Akiyama, Y., Kihara, A., Tokuda, H., and Ito, K.: "FtsH (HflB) is an ATP-dependent protease selectively acting on SecY and some other membrane proteins." J.Biol. Chem.271. 31196-31201 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Homma, T., Yoshihisa, T., Kihara, A., Akiyama, Y., and Ito, K.: "Intracellular stability of a fragment of beta-galactosidase : effects of amino-terminally fused polypeptides." Biochem. Biophys. Res. Commun.215. 452-458 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Watanabe, M., and Biobel, G.: "High-affinity binding of Escherichia coli SecB to the signal sequence region of a presecretory protein" Proc. Natl. Acad. Sci. USA. 92. 10133-10136 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Akiyama, Y., Kihara, A., and Ito, K.: "Subunit a of proton ATPase F_0 sector is a substrate of the FtsH protease in Escherichia coli." FEBS Lett.399. 26-28 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kishigami, S., and Ito, K.: "Roles of cysteine residues of DsbB in its activity to reoxidize DsbA,the protein disulfide bond catalyst of Escherichia coli." Genes to Cells. 1. 201-208 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Sone, M., Kishigami, S., Yoshihisa, T., and Ito, K.: "Roles of disulfide bonds in bacterial alkaline phosphatase." J.Biol. Chem.(in press). (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Endo, T., Mitsui, S., and Roise, D.: "Mitochondrial presequences can induce aggregation of unfolded proteins." FEBS Lett.359. 93-96 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nohara, T., Nakai, M., Goto, A., and Endo, T.: "Isolation and characterization of the cDNA for pea chloroplast SecA : evolutionary conservation of the bacterial-type SecA-dependent protein transport within chloroplasts." FEBS Lett.364. 305-308 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Endo, T., Nakayama, Y., and Nakai, M.: "Avidin fusion protein as a tool to genecrate a stable translocation intermediate spanning the mitochondrial membranes." J.Biochem.118. 753-759 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Endo, T., Mitsui, S., Nakai, M., and Roise, D.: "Binding of mithochondrial presequences to yeast cytosolic heat shock protein 70 depends of the amphiphilicity of the presequence." J.Bio. Chem.271. 4161-4167 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kanamori, T., Nishikawa, S., Shin, I., Schultz, P.G., and Endo, T.: "Probing the environment along the protein import pathways in yeast mitochondria by site-specific photocrosslinking." Proc. Natl. Acad. Sci. U.S.A.94. (in press). (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Murai, N., Kamata, H., Nagashima, Y., Yagisawa, H., and Hirata, H.: "A Novel Insertion Sequence (IS)-like Element of the Thermophilic Bacterium PS3 promotes Expression of the Alanine Carrier Protein-encoding Gene." Gene. 163. 103-107 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Jin, T., Kinjo, M., Koyama, T., Kobayashi, Y.and Hirata, H.: "Selective Na_+ Transport through Phospholipid Bilayr Membrane by a Synthetic Calix [4] arene Carrier." Langmuir. 12. 2684-2689 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kamata, H., Tanaka, C., Yagisawa, H.and Hirata, H.: "Nerve Growth Factor and Forskolin prevent H_2O_2-induced Apoptosis in PC12 Cells by Glutathione Independent Mechanism." Neurosci. Lett.212. 179-182 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kamata, H., Tanaka, C., Yagisawa, H., Matsuda, S., Gotoh, Y., Nishida, E.and Hirata, H.: "Involvement of Cellular Redox State in Cell Survival and Neuronal Differentiation in PC12 Cells." Biochem. Soc. Trans.24. 541 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kamata, H., Tanaka, C., Yagisawa, H., Matsuda, S., Gotoh, Y., Nishida, E.and Hirata, H.: "Suppression on NGF-induced Neuronal Differentiation of PC12 Cells ; N-Acetylcysteine (NAC) uncouples the Signal Transduction from Ras to the MAP Kinase Cascade." J.Biol. Chem.271. 33018-33025 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Uchida, K., Mori, H., and Mizushima S.: "Stepwise Movement of Preproteins in the Process of Translocation across the Cytoplasmic Membrane of Escherichia coli." J.Biol. Chem.270. 30862-30868 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Sato, K., Mori, H., Yoshida, M., and Mizushima, S.: "Characterization of a Potential Catalytic Residue, Asp-133, in the High Affinity ATP-binding Site of Escherichia coli SecA,Translocation ATPase." J.Biol. Chem.271. 17439-17444 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Sato, K., Mori, H., Yoshida, M., Tagaya, M., and Mizushima, S.: "Short hydrophobic segments in the mature domain of proompA determine its stepwise movement during translocation across the cytoplasmic membrane of Escherichia coli" J.Biol. Chem.272 (in press). (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nakane, A., Takamatsu, H., Oguro, A., Sadaie, Y., Nakamura, K., and Yamane, K.: "Acquisition of azide-resistance by clevated SecA ATPase activity confers azide-resistance upon cell growth and protein translocation in Bacillus subtilis." Microbiology. 141. 113-121 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ogawa, K., Akagawa, E., Yamane, K., Z.-W.Sun, M.LaCelle, P.Zuber and Nakano, M.M.: "The nasB operon and nasA gene are required for nitrate/nitrite assimilation in Bacillus subtilis." J.Bacteriol.177. 1409-1413 (1995)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kurita, K., Honda, K., Suzuma, S., Takamatsu, H., Nakamura, K., and Yamane, K.: "Identification of a region of Bacillus subtilis Ffh, a homologue of mammalian SRP54 protein, that is essential for binding to small cytoplasmic RNA." J.Biol. Chem.271. 13140-13146 (1996)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Yamane, K., Kumano, M., and Kurita, K.: "25^0-36^0 region of the Bacillus subtilis chromomose : determination of the sequence of a 146 kb segment and identification of 113 genes." Microbiology. 142. 3047-3056 (1996)
  • Description: 「研究成果報告書概要(欧文)」より