Thermostable chemotaxis proteins from the hyperthermophilic bacterium Thermotoga maritima
10.1128/jb.178.2.484-489.1996
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

An expressed sequence tag homologous to cheA was previously isolated by random sequencing of Thermotoga maritima cDNA clones (C. W. Kim, P. Markiewicz, J. J. Lee, C. F. Schierle, and J. H. Miller, J. Mol. Biol. 231: 960-981, 1993). Oligonucleotides complementary to this sequence tag were synthesized and used to identify a clone from a T. maritima lambda library by using PCR. Two partially overlapping restriction fragments were subcloned from the lambda clone and sequenced. The resulting 5,251-bp sequence contained five open reading frames, including cheA, cheW, and cheY. In addition to the chemotaxis genes, the fragment also encodes a putative protein isoaspartyl methyltransferase and an open reading frame of unknown function. Both the cheW and cheY genes were individually cloned into inducible Escherichia coli expression vectors. Upon induction, both proteins were synthesized at high levels. T. maritima CheW and CheY were both soluble and were easily purified from the bulk of the endogenous E. coli protein by heat treatment at 80 degrees C for 10 min. CheY prepared in this way was shown to be active by the demonstration of Mg(2+)-dependent autophosphorylation with [32P]acetyl phosphate. In E. coli, CheW mediates the physical coupling of the receptors to the kinase CheA. The availability of a thermostable homolog of CheW opens the possibility of structural characterization of this small coupling protein, which is among the least well characterized proteins in the bacterial chemotaxis signal transduction pathway.

Bibliography

Swanson, R. V., Sanna, M. G., & Simon, M. I. (1996). Thermostable chemotaxis proteins from the hyperthermophilic bacterium Thermotoga maritima. Journal of Bacteriology, 178(2), 484–489.

Authors 3
  1. R V Swanson (first)
  2. M G Sanna (additional)
  3. M I Simon (additional)
References 40 Referenced 42
  1. Altschul , S. F. , W. Gish , W. Miller , E. W. Myers , and D. J. Lipman . 1990 . Basic local alignment search tool . J. Mol. Biol. 215 : 403 - 410 . (10.1016/S0022-2836(05)80360-2) / J. Mol. Biol. / Basic local alignment search tool by Altschul S. F. (1990)
  2. Bischoff , D. S. , R. B. Bourret , M. L. Kirsch , and G. W. Ordal . 1993 . Purification and characterization of Bacillus subtilis CheY . Biochemistry 32 : 9256 - 9261 . (10.1021/bi00086a035) / Biochemistry / Purification and characterization of Bacillus subtilis CheY by Bischoff D. S. (1993)
  3. Bischoff , D. S. , and G. W. Ordal . 1991 . Sequence and characterization of Bacillus subtilis CheB, a homolog of Escherichia coli CheY, and its role in a different mechanism of chemotaxis . J. Biol. Chem. 266 : 12301 - 12305 . (10.1016/S0021-9258(18)98896-4) / J. Biol. Chem. / Sequence and characterization of Bacillus subtilis CheB, a homolog of Escherichia coli CheY, and its role in a different mechanism of chemotaxis by Bischoff D. S. (1991)
  4. Bischoff , D. S. , and G. W. Ordal . 1992 . Bacillus subtilis chemotaxis: a deviation from the Escherichia paradigm . Mol. Microbiol. 6 : 23 - 28 . (10.1111/j.1365-2958.1992.tb00833.x) / Mol. Microbiol. / Bacillus subtilis chemotaxis: a deviation from the Escherichia paradigm by Bischoff D. S. (1992)
  5. Blamey , J. M. , and M. W. W. Adams . 1994 . Characterization of an ancestral type of pyruvate ferredoxin oxidoreductase from the hyperthermophilic bacterium, Thermotoga maritima . Biochemistry 33 : 1000 - 1007 . (10.1021/bi00170a019) / Biochemistry / Characterization of an ancestral type of pyruvate ferredoxin oxidoreductase from the hyperthermophilic bacterium, Thermotoga maritima by Blamey J. M. (1994)
  6. Borkovich , K. A. , N. Kaplan , J. F. Hess , and M. I. Simon . 1989 . Transmembrane signal transduction in bacterial chemotaxis involves ligand-dependent activation of phosphate group transfer . Proc. Natl. Acad. Sci. USA 86 : 1208 - 1212 . (10.1073/pnas.86.4.1208) / Proc. Natl. Acad. Sci. USA / Transmembrane signal transduction in bacterial chemotaxis involves ligand-dependent activation of phosphate group transfer by Borkovich K. A. (1989)
  7. Bourret , R. B. , J. Davagnino , and M. I. Simon . 1993 . The carboxy-terminal portion of the CheA kinase mediates regulation of autophosphorylation by transducer and CheW . J. Bacteriol. 175 : 2097 - 2101 . (10.1128/jb.175.7.2097-2101.1993) / J. Bacteriol. / The carboxy-terminal portion of the CheA kinase mediates regulation of autophosphorylation by transducer and CheW by Bourret R. B. (1993)
  8. Bourret , R. B. , J. F. Hess , and M. I. Simon . 1990 . Conserved aspartate residues and phosphorylation in signal transduction by the chemotaxis protein CheY . Proc. Natl. Acad. Sci. USA 87 : 41 - 45 . (10.1073/pnas.87.1.41) / Proc. Natl. Acad. Sci. USA / Conserved aspartate residues and phosphorylation in signal transduction by the chemotaxis protein CheY by Bourret R. B. (1990)
  9. Brennan , T. V. , J. W. Anderson , Z. Jia , E. B. Waygood , and S. Clarke . 1994 . Repair of spontaneously deamidated Hpr phosphocarrier protein catalyzed by the L-isoaspartyl-(D-aspartate) O-methyltransferase . J. Biol. Chem. 269 : 24586 - 24595 . (10.1016/S0021-9258(17)31432-1) / J. Biol. Chem. / Repair of spontaneously deamidated Hpr phosphocarrier protein catalyzed by the L-isoaspartyl-(D-aspartate) O-methyltransferase by Brennan T. V. (1994)
  10. Bronnenmeir , K. , A. Kern , W. Liebl , and W. L. Staudenbauer . 1995 . Purification of Thermotoga maritima enzymes for the degradation of cellulosic materials . Appl. Environ. Microbiol. 61 : 1399 - 1407 . (10.1128/AEM.61.4.1399-1407.1995) / Appl. Environ. Microbiol. / Purification of Thermotoga maritima enzymes for the degradation of cellulosic materials by Bronnenmeir K. (1995)
  11. Brown , S. H. , C. Sjoholm , and R. M. Kelly . 1993 . Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima . Biotechnol. Bioeng. 41 : 878 - 886 . (10.1002/bit.260410907) / Biotechnol. Bioeng. / Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima by Brown S. H. (1993)
  12. Cohen , S. P. , H. Haechler , and S. B. Levy . 1993 . Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli . J. Bacteriol. 175 : 1484 - 1492 . (10.1128/jb.175.5.1484-1492.1993) / J. Bacteriol. / Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli by Cohen S. P. (1993)
  13. Fuhrer , D. K. , and G. W. Ordal . 1991 . Bacillus subtilis CheN, a homolog of CheA, the central regulator of chemotaxis in Escherichia coli . J. Bacteriol. 173 : 7443 - 7448 . (10.1128/JB.173.23.7443-7448.1991) / J. Bacteriol. / Bacillus subtilis CheN, a homolog of CheA, the central regulator of chemotaxis in Escherichia coli by Fuhrer D. K. (1991)
  14. Gabelsberger , J. , W. Liebel , and K. H. Schleifer . 1993 . Purification and properties of recombinant ~-glucosidase of the hyperthermophilic bacterium Thermotoga maritima . Appl. Microbiol. Biotechnol. 40 : 44 - 52 . (10.1007/BF00170427) / Appl. Microbiol. Biotechnol. / Purification and properties of recombinant ~-glucosidase of the hyperthermophilic bacterium Thermotoga maritima by Gabelsberger J. (1993)
  15. Gegner , J. A. , D. R. Graham , A. F. Roth , and F. W. Dahlquist . 1992 . Assembly of an MCP receptor, CheW, and kinase CheA complex in the bacterial chemotaxis signal transduction pathway . Cell 70 : 975 - 982 . (10.1016/0092-8674(92)90247-A) / Cell / Assembly of an MCP receptor, CheW, and kinase CheA complex in the bacterial chemotaxis signal transduction pathway by Gegner J. A. (1992)
  16. Greck , M. , J. Platzer , V. Sourjik , and R. Schmidt . 1995 . Analysis of a chemotaxis operon in Rhizobium melliloti . Mol. Microbiol. 15 : 989 - 1000 . (10.1111/j.1365-2958.1995.tb02274.x) / Mol. Microbiol. / Analysis of a chemotaxis operon in Rhizobium melliloti by Greck M. (1995)
  17. Hess , J. F. , R. B. Bourret , and M. I. Simon . 1988 . Histidine phosphorylation and phosphoryl group transfer in bacterial chemotaxis . Nature (London) 336 : 139 - 143 . (10.1038/336139a0) / Nature (London) / Histidine phosphorylation and phosphoryl group transfer in bacterial chemotaxis by Hess J. F. (1988)
  18. Huber , R. , T. A. Langworthy , H. Konig , M. Thomm , C. R. Woese , U. B. Sleyter , and K. O. Stetter . 1986 . Thermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up to 90~C . Arch. Microbiol. 144 : 324 - 333 . (10.1007/BF00409880) / Arch. Microbiol. / Thermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up to 90~C by Huber R. (1986)
  19. Kim , C. W. , P. Markiewicz , J. J. Lee , C. F. Schierle , and J. H. Miller . 1993 . Studies of the hyperthermophile Thermotoga maritima by random sequencing of cDNA and genomic libraries . J. Mol. Biol. 231 : 960 - 981 . (10.1006/jmbi.1993.1345) / J. Mol. Biol. / Studies of the hyperthermophile Thermotoga maritima by random sequencing of cDNA and genomic libraries by Kim C. W. (1993)
  20. Kofoid , E. C. , and J. S. Parkinson . 1991 . Tandem translation starts in the cheA locus of Escherichia coli . J. Bacteriol. 173 : 2116 - 2119 . (10.1128/JB.173.6.2116-2119.1991) / J. Bacteriol. / Tandem translation starts in the cheA locus of Escherichia coli by Kofoid E. C. (1991)
  21. Li , J. , R. V. Swanson , M. I. Simon , and R. M. Weis . 1995 . The response regulators CheB and CheY exhibit competitive binding to the kinase CheA . Biochemistry 34 : 14626 - 14636 . (10.1021/bi00045a003) / Biochemistry / The response regulators CheB and CheY exhibit competitive binding to the kinase CheA by Li J. (1995)
  22. Liebel , W. , R. Feil , J. Gabelsberger , J. Kellermann , and K. H. Schleifer . 1993 . Purification and characterization of a novel thermostable 4-~-glucanotransferase of Thermotoga maritima cloned in Escherichia coli . Eur. J. Biochem. 207 : 81 - 88 . (10.1111/j.1432-1033.1992.tb17023.x) / Eur. J. Biochem. / Purification and characterization of a novel thermostable 4-~-glucanotransferase of Thermotoga maritima cloned in Escherichia coli by Liebel W. (1993)
  23. Lowry , D. L. , A. F. Roth , A , P. B. Rupert , F. W. Dahlquist , F. J. Moy , P. J. Domaille , and P. Matsumura . 1994 . Signal-transduction in chemotaxis, a propagating conformation change upon phosphorylation of CheY . J. Biol. Chem. 269 : 26358 - 26362 . (10.1016/S0021-9258(18)47201-8) / J. Biol. Chem. / Signal-transduction in chemotaxis, a propagating conformation change upon phosphorylation of CheY by Lowry D. L. (1994)
  24. Lukat , G. S. , W. R. McCleary , A. M. Stock , and J. B. Stock . 1992 . Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors . Proc. Natl. Acad. Sci. USA 89 : 718 - 722 . (10.1073/pnas.89.2.718) / Proc. Natl. Acad. Sci. USA / Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors by Lukat G. S. (1992)
  25. Morrison , T. B. , and J. S. Parkinson . 1994 . Liberation of an interaction domain from the phosphotransfer region of CheA, a signaling kinase of Escherichia-coli . Proc. Natl. Acad. Sci. USA 91 : 5485 - 5489 . (10.1073/pnas.91.12.5485) / Proc. Natl. Acad. Sci. USA / Liberation of an interaction domain from the phosphotransfer region of CheA, a signaling kinase of Escherichia-coli by Morrison T. B. (1994)
  26. Moy , F. J. , D. F. Lowry , P. Matsumura , F. W. Dahlquist , J. E. Krywko , and P. J. Domaille . 1994 . Assignments secondary structure, global fold, and dynamics of chemotaxis Y protein using three- and four-dimensional heteronuclear (13C, 15N) NMR spectroscopy . Biochemistry 33 : 10731 - 10742 . (10.1021/bi00201a022) / Biochemistry / Assignments secondary structure, global fold, and dynamics of chemotaxis Y protein using three- and four-dimensional heteronuclear (13C, 15N) NMR spectroscopy by Moy F. J. (1994)
  27. Parkinson , J. S. , and E. C. Kofoid . 1993 . Communication modules in bacterial signaling proteins. Annu . Rev. Genet. 26 : 71 - 112 . (10.1146/annurev.ge.26.120192.000443) / Rev. Genet. / Communication modules in bacterial signaling proteins. Annu by Parkinson J. S. (1993)
  28. Saiki , R. K. , D. H. Gelfand , S. Stoffel , S. J. Scharf , R. Higuchi , G. T. Horn , K. B. Mullis , and H. A. Erlich . 1988 . Primer-directed enzymatic amplification of DNA with a thermostable DNA-polymerase . Science 239 : 487 - 489 . (10.1126/science.239.4839.487) / Science / Primer-directed enzymatic amplification of DNA with a thermostable DNA-polymerase by Saiki R. K. (1988)
  29. Sanna , M. G. , R. V. Swanson , R. B. Bourret , and M. I. Simon . 1995 . Mutations in the chemotactic response regulator CheY that confer resistance to the phosphatase activity of CheZ . Mol. Microbiol. 15 : 1069 - 1079 . (10.1111/j.1365-2958.1995.tb02282.x) / Mol. Microbiol. / Mutations in the chemotactic response regulator CheY that confer resistance to the phosphatase activity of CheZ by Sanna M. G. (1995)
  30. Schuster , S. C. , R. V. Swanson , L. A. Alex , R. B. Bourret , and M. I. Simon . 1993 . Assembly and function of a signal transduction complex monitored by surface plasmon resonance . Nature (London) 365 : 343 - 347 . (10.1038/365343a0) / Nature (London) / Assembly and function of a signal transduction complex monitored by surface plasmon resonance by Schuster S. C. (1993)
  31. Stock , A. , D. E. Koshland , and J. Stock . 1985 . Homologies between the Salmonella typhimurium CheY protein and proteins involved in the regulation of chemotaxis, membrane protein synthesis, and sporulation . Proc. Natl. Acad. Sci. USA 82 : 7989 - 7993 . (10.1073/pnas.82.23.7989) / Proc. Natl. Acad. Sci. USA / Homologies between the Salmonella typhimurium CheY protein and proteins involved in the regulation of chemotaxis, membrane protein synthesis, and sporulation by Stock A. (1985)
  32. Stock , A. M. , E. Martinez-Hackert , B. F. Rasmussen , A. West , J. B. Stock , D. Ringe , and G. A. Petsko . 1993 . Structure of the Mg2~ bound form of CheY and mechanism of phosphoryl transfer in bacterial chemotaxis . Biochemistry 32 : 13376 - 13380 . (10.1021/bi00212a001) / Biochemistry / Structure of the Mg2~ bound form of CheY and mechanism of phosphoryl transfer in bacterial chemotaxis by Stock A. M. (1993)
  33. Swanson , R. V. , D. F. Lowry , P. Matsumura , M. McEvoy , M. I. Simon , and F. W. Dahlquist . 1995 . Localized perturbations in CheY structure monitored by NMR identify a CheA binding interface . Nature Struct. Biol. 2 : 906 - 910 . (10.1038/nsb1095-906) / Nature Struct. Biol. / Localized perturbations in CheY structure monitored by NMR identify a CheA binding interface by Swanson R. V. (1995)
  34. Swanson , R. V. , S. C. Schuster , and M. I. Simon . 1993 . Expression of CheA fragments which define domains encoding kinase, phosphotransfer, and CheY binding activities . Biochemistry 32 : 7623 - 7629 . (10.1021/bi00081a004) / Biochemistry / Expression of CheA fragments which define domains encoding kinase, phosphotransfer, and CheY binding activities by Swanson R. V. (1993)
  35. Volkman , B. F. , M. J. Nohaile , N. K. Amy , S. Kustu , and D. E. Wemmer . 1995 . 3-dimensional solution structure of the N-terminal receiver domain of NtrC . Biochemistry 34 : 1413 - 1424 . (10.1021/bi00004a036) / Biochemistry / 3-dimensional solution structure of the N-terminal receiver domain of NtrC by Volkman B. F. (1995)
  36. Volz , K. 1993 . Structural conservation in the CheY superfamily . Biochemistry 32 : 11741 - 11753 . (10.1021/bi00095a001) / Biochemistry / Structural conservation in the CheY superfamily by Volz K. (1993)
  37. Wrba , A. , R. Jaenicke , R. Huber , and K. O. Stetter . 1990 . Lactate dehydrogenase from the extreme thermophile Thermotoga maritima . Eur. J. Biochem. 188 : 195 - 201 . (10.1111/j.1432-1033.1990.tb15388.x) / Eur. J. Biochem. / Lactate dehydrogenase from the extreme thermophile Thermotoga maritima by Wrba A. (1990)
  38. Wrba , A. , A. Schweiger , V. Schultes , and R. Jaenicke . 1990 . Extremely thermostable D-glyceraldehyde-3-phosphate dehydrogenase from the eubacterium Thermotoga maritima . Biochemistry 29 : 7584 - 7592 . (10.1021/bi00485a007) / Biochemistry / Extremely thermostable D-glyceraldehyde-3-phosphate dehydrogenase from the eubacterium Thermotoga maritima by Wrba A. (1990)
  39. Zamenhof , P. J. , and M. Villarejo . 1972 . Construction and properties of Escherichia coli strains exhibiting ~-complementation of ~-galactosidase fragments in vivo . J. Bacteriol. 110 : 171 - 178 . (10.1128/jb.110.1.171-178.1972) / J. Bacteriol. / Construction and properties of Escherichia coli strains exhibiting ~-complementation of ~-galactosidase fragments in vivo by Zamenhof P. J. (1972)
  40. Zusman , D. R. , and M. J. McBride . 1991 . Sensory transduction in the gliding bacterium Myxococcus xanthus . Mol. Microbiol. 5 : 2323 - 2329 . (10.1111/j.1365-2958.1991.tb02077.x) / Mol. Microbiol. / Sensory transduction in the gliding bacterium Myxococcus xanthus by Zusman D. R. (1991)
Dates
Type When
Created 8 years, 9 months ago (Nov. 10, 2016, 8:22 a.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:59 p.m.)
Indexed 1 year, 10 months ago (Oct. 28, 2023, 8:42 p.m.)
Issued 29 years, 8 months ago (Jan. 1, 1996)
Published 29 years, 8 months ago (Jan. 1, 1996)
Published Print 29 years, 8 months ago (Jan. 1, 1996)
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@article{Swanson_1996, title={Thermostable chemotaxis proteins from the hyperthermophilic bacterium Thermotoga maritima}, volume={178}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.178.2.484-489.1996}, DOI={10.1128/jb.178.2.484-489.1996}, number={2}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Swanson, R V and Sanna, M G and Simon, M I}, year={1996}, month=jan, pages={484–489} }