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Masayori Inouye
Research Fields
Research DescriptionOur laboratory is interested in molecular mechanisms of cellular adaptation to various stresses and growth conditions. Cells contain a large number of sensory proteins responding to different stresses. Stress signals are transduced by these sensory proteins across the membrane to regulate expression of a specific group of genes essential for adaptation. In prokaryotes, protein histidine kinases play the major role as sensory proteins. Recently, we determined the three-dimensional structure of a histidine kinase of E. coli functioning as an osmosensor. Biochemical, biophysical and genetic approaches are used to further elucidate how this kinase tranduces signals and activates its downstream transcription factor. We are also interested in cellular adaptation to low temperature. In E. coli, a specific group of so called "cold-shock" proteins are induced. Interestingly, E. coli contains a large family of CspA, the major cold-shock protein, which is also highly conserved to human. We investigate how cold-shock genes are specifically induced at low temperature and what are the function of cold-shock proteins. We also study cold-shock response in human cells. Protein folding is also our research interest. We have shown that a group of proteins require an extra polypeptide called intramolecular chaperones (IMC) for their folding. Mutations in IMC block protein folding pathways. Since IMC is not a part of the final folded protein, the IMC-mediated protein folding is an ideal system for studying how protein structures are determined on the basis of their primary sequences. Recently, we demonstrated a phenomenon called protein memory, where an identical polypeptide can be folded into two different structures having different enzymatic specificities. In addition, we are interested in cell division regulated by a GTP-binding protein in E. coli. This protein called Era (E. coli Ras-like protein) is likely to be involved in an early stage of septum formation. We also investigate a bacterial retroelement responsible for the synthesis of a unique single-stranded DNA. This system was originally discovered in our laboratory and we attempt to elucidate how bacterial reverse transcriptase are able to initiate cDNA synthesis from a 2'OH group of an internal G residue in a template RNA molecule. This study also provides insights into the origin of reverse transcriptases and retroelements. Selected PublicationsI. Protein Histidine Kinase-OsmoregulationUtsumi, R., R.E. Brissette, A. Rampersaud, S.A. Forst, K. Oosawa, and M. Inouye (1989) Activation of Bacterial Porin Gene Expression by a Chimeric Signal Transducer in Response to Aspartate. Science 245: 1246 1249. Yang, Y., and M. Inouye (1991) Intermolecular Complementation between Two Defective Mutants of Escherichia coli Signal transducing Receptors. Proc. Natl. Acad. Sci. USA 88: 11057 11061. Yang, Y. and M. Inouye (1993) Requirement of Both Kinase and Phosphatase Activites of an Escherichia coli Receptor (Taz1) for Ligand dependent Signal Transduction. J. Mol. Biol. 231: 335 342. Yang, Y., H. Park and M. Inouye (1993) Ligand Binding Induces Asymmetrical Transmembrane Signal through a Receptor Dimer. J. Mol. Biol. 232: 493 498. Rampersaud, A., S. Harlocker, and M. Inouye (1994) The OmpR Protein of Escherichia coli Binds to Sites in the ompF Promoter Region in a Hierarchical Manner Determined by Its Degree of Phosphorylation. J. Biol. Chem. 269: 12559 12566. Harlocker, S., L. Bergstrom, and M. Inouye (1995) Tandem
Binding of Six OmpR Proteins to the ompF Upstream Regulatory Sequence
of Escherichia coli. J. Biol. Chem. 270: 26849-26856. Egger, L. A., H. Park and M. Inouye (1997) Review: Signal Transduction via the Histidyl-aspartyl Phosphorelay. Genes to Cells 2: 167-18 II. Cold-Shock ResponseGoldstein, J., N.S. Pollitt, and M. Inouye (1990) The Major Cold Shock Protein of Escherichia coli; Transcriptional Regulation of its Gene. Proc. Natl. Acad. Sci. USA 87: 283 287. Newkirk, K., W. Feng, W. Jiang, R. Tejero, S.D. Emerson, M. Inouye, G. Montelione (1994) Solution NMR Structure of the Major Cold shock Proteins (CspA) from Escherichia coli: Identification of a Binding Epitope for DNA. Proc. Natl. Acad. Sci. USA 91: 5114 5118. Schindelin, H., F. Cordes, W. Jiang, M. Inouye, and U. Heinemann (1994) Crystal Structure of the Major Cold shock Protein of Escherichia coli. Proc. Natl. Acad. Sci. USA 91: 5119 5123. Jones, P. and M. Inouye (1996) RbfA, a 30S-Ribosomal Binding Factor, is a Cold-shock Protein Whose Absence Triggers the Cold-shock Response. Molec. Micro. 21: 1207-1218. Jiang, W., Y. Hou, and M. Inouye (1997) CspA, The Major Cold-shock Protein of Escherichia coli, is an RNA Binding Protein. J. Biol. Chem. 272: 196-202. Mitta, M., L. Fang, and M. Inouye (1997) Deletion Analysis of cspA of Escherichia coli: Requirement for the AT-rich UP Element for cspA Transcription and the Downstream Box in the coding Region for its Cold Shock Induction. Molec. Micro. 26: 321-335. Bae, W., P.G. Jones, and M. Inouye (1997) CspA, the Major Cold-shock Protein of Escherichia coli, Negatively Regulates Its Own Gene Expression. J. Bacteriol. 179:7081-7088. Fang, L., Y. Hou, and M. Inouye (1998) Role of the Cold-Box Region in the 5' Untranslated Region of the cspA mRNA for Its Transient Expression at Low Temperature in Escherichia coli. J. Bacteriol. 180(1):90-95. Yamanaka, K., Fang, L., and Inouye, M. (1998). The CspA family of Escherichia coli: multiple gene duplication for stress adaptation. Molec. Microbio. 27(2):247-256. III. Intramolecular Chaperone and Protein FoldingIkemura, H., H. Takagi, and M. Inouye (1987) Requirement of Pro sequence for the Production of Active Subtilisin E in Escherichia coli. J. Biol. Chem. 262: 7859 7864. Zhu, X., Y. Ohta, F. Jordan, and M. Inouye (1989) Pro
sequence of Subtilisin Can Guide the Refolding of Denatured Subtilisin
in an Intermolecular Process. Nature 339: 483 484. Li, Y. and M. Inouye (1994) Autoprocessing of Prothiolsubtilisin E in Which the Active Site Serine 221 is Altered to Cysteine. J. Biol. Chem. 269: 4169 4174. Shinde, U. and M. Inouye (1995) Folding Mediated by an Intramolecular Chaperone: Autoprocessing Pathway of the Precursor Resolved Via a `Substrate assisted Catalysis' Mechanism. J. Mol. Biol. 247: 390 395. Shinde, U. and M. Inouye (1995) Folding Pathway Mediated by an Intramolecular Chaperone: Characterization of the Structural Changes in Pro subtilisin E Coincident with Autoprocessing. J. Mol. Biol. 252: 25-30. Shinde, U.P., J.J. Liu, and M. Inouye (1997) Protein Memory through Altered Folding Mediated by Intramolecular Chaperones. Nature 398: 520-522. IV. Other Selected PublicationsMizuno, T., M. Y. Chou, and M. Inouye (1984) A Unique Mechanism Regulating Gene Expression: Translational Inhibition by a Complementary RNA Transcript (micRNA). Proc. Natl. Acad. Sci. USA 81:1966 1970. Yee, T., T. Furuichi, S. Inouye, and M. Inouye (1984) Multicopy Single Stranded DNA Isolated from a Gram Negative Bacterium, Myococcus xanthus. Cell 38:203 209. Coleman, J., P. J. Green and M. Inouye (1984) The Use of RNAs Complementary to Specific mRNAs to Regulate the Expression of Individual Bacterial Genes. Cell 37:429 436. Coleman, J., A. Hirashima, Y. Inokuchi, P.J. Green, and M. Inouye (1985) A Novel Immune System Against Bacteriophage Infection Using Complementary RNA (micRNA). Nature 315:601 603. Furuichi, T., S. Inouye, and M. Inouye (1987) Biosynthesis and Structure of Stable Branched RNA Covalently Linked to the 5' End of Multicopy Single Stranded DNA of Stigmatella aurantiaca. Cell 48:55 62. Yamaguchi, K., F. Yu, M. Inouye (1988) A Single Amino Acid Determinant of the Membrane Localization of Lipoproteins in E. coli. Cell 53: 423 432. Lampson, B.C., J. Sun, M. Y. Hsu, J. Vallejo Ramirez, S. Inouye and M. Inouye (1989) Reverse Transcriptase in a Clinical Strain of Escherichia coli: Production of Branched RNA linked msDNA. Science 243: 1033 1038. Inouye, S., M. Y. Hsu, S. Eagle and M. Inouye (1989) Reverse Transcriptase Associated with the Biosynthesis of the Branched RNA linked msDNA in Myococcus xanthus. Cell 56: 709 717. Lerner, C.G. and M. Inouye (1991) Pleiotropic Effects of Depletion of ERA, an Essential GTP binding Protein in E. coli. Mol. Micro. 5: 951 957. Munoz Dorado, J., S. Inouye, and M. Inouye (1991) A Gene Encoding a Protein Serine/Threonine Kinase is Required for Normal Development of Myxococcus xanthus, a Gram negative Bacterium. Cell 67: 995 1006. Mirochnitchenko, O, U. Palnitkar, M. Philbert and M. Inouye (1995) Thermosensitive Phenotype of Transgenic Mice Overproducing Human Glutathione Peroxidases. Proc. Natl. Acad. Sci. USA 92: 8120-8124. Mao, J.-R., M. Shimada, S. Inouye, and M. Inouye (1995) Gene Regulation by Antisense DNA Produced in Vivo. J. Biol. Chem. 270: 19684-19687. Inouye LaboratoryGraduate Students:
Postdoctoral Researchers:
Staff:
Adjunct Assistant Professors:
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