Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1996 Jan 2;15(1):162–171.

Involvement of stress protein PspA (phage shock protein A) of Escherichia coli in maintenance of the protonmotive force under stress conditions.

M Kleerebezem 1, W Crielaard 1, J Tommassen 1
PMCID: PMC449928  PMID: 8598199

Abstract

The expression of specific PhoE mutant proteins leads to induction of the expression of the psp operon of Escherichia coli and the export of various plasmid-encoded precursors is retarded in a pspA mutant strain. Here, we have investigated the specific role of various Psp proteins in the export process. PspB and PspC are both inner membrane proteins that are involved in the regulation of the transcription of the psp operon. Precursor PhoE translocation was retarded in a pspB mutant strain to a similar extent as in a pspA mutant strain. The reduced translocation efficiencies in the various psp mutants could be complemented by expression of PspA from a plasmid, indicating that only PspA is required for efficient translocation. Mutant prePhoE proteins that can be translocated independently of the deltamu H+ appeared to translocate equally efficiently in a wild-type and in a pspA mutant strain. Furthermore, quantitative in vivo determination of the deltamu H+ showed that it specifically decreased in a pspA mutant strain upon expression of plasmid-encoded (mutant) prePhoE protein. Apparently, the translocation defects observed in a psp mutant strain are caused by a decrease of the delta mu H+ and PspA functions by maintaining the delta mu H+ under these conditions.

Full text

PDF
162

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agterberg M., Adriaanse H., van Bruggen A., Karperien M., Tommassen J. Outer-membrane PhoE protein of Escherichia coli K-12 as an exposure vector: possibilities and limitations. Gene. 1990 Mar 30;88(1):37–45. doi: 10.1016/0378-1119(90)90057-x. [DOI] [PubMed] [Google Scholar]
  2. Arkowitz R. A., Wickner W. SecD and SecF are required for the proton electrochemical gradient stimulation of preprotein translocation. EMBO J. 1994 Feb 15;13(4):954–963. doi: 10.1002/j.1460-2075.1994.tb06340.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bassford P. J., Jr, Silhavy T. J., Beckwith J. R. Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm. J Bacteriol. 1979 Jul;139(1):19–31. doi: 10.1128/jb.139.1.19-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bieker K. L., Silhavy T. J. PrlA (SecY) and PrlG (SecE) interact directly and function sequentially during protein translocation in E. coli. Cell. 1990 Jun 1;61(5):833–842. doi: 10.1016/0092-8674(90)90193-i. [DOI] [PubMed] [Google Scholar]
  5. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bosch D., Leunissen J., Verbakel J., de Jong M., van Erp H., Tommassen J. Periplasmic accumulation of truncated forms of outer-membrane PhoE protein of Escherichia coli K-12. J Mol Biol. 1986 Jun 5;189(3):449–455. doi: 10.1016/0022-2836(86)90316-5. [DOI] [PubMed] [Google Scholar]
  7. Bosch D., de Boer P., Bitter W., Tommassen J. The role of the positively charged N-terminus of the signal sequence of E. coli outer membrane protein PhoE in export. Biochim Biophys Acta. 1989 Feb 13;979(1):69–76. doi: 10.1016/0005-2736(89)90524-5. [DOI] [PubMed] [Google Scholar]
  8. Brissette J. L., Russel M., Weiner L., Model P. Phage shock protein, a stress protein of Escherichia coli. Proc Natl Acad Sci U S A. 1990 Feb;87(3):862–866. doi: 10.1073/pnas.87.3.862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brissette J. L., Weiner L., Ripmaster T. L., Model P. Characterization and sequence of the Escherichia coli stress-induced psp operon. J Mol Biol. 1991 Jul 5;220(1):35–48. doi: 10.1016/0022-2836(91)90379-k. [DOI] [PubMed] [Google Scholar]
  10. Brundage L., Hendrick J. P., Schiebel E., Driessen A. J., Wickner W. The purified E. coli integral membrane protein SecY/E is sufficient for reconstitution of SecA-dependent precursor protein translocation. Cell. 1990 Aug 24;62(4):649–657. doi: 10.1016/0092-8674(90)90111-q. [DOI] [PubMed] [Google Scholar]
  11. Carlson J. H., Silhavy T. J. Signal sequence processing is required for the assembly of LamB trimers in the outer membrane of Escherichia coli. J Bacteriol. 1993 Jun;175(11):3327–3334. doi: 10.1128/jb.175.11.3327-3334.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chun S. Y., Randall L. L. In vivo studies of the role of SecA during protein export in Escherichia coli. J Bacteriol. 1994 Jul;176(14):4197–4203. doi: 10.1128/jb.176.14.4197-4203.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. De Vrije T., Tommassen J., De Kruijff B. Optimal posttranslational translocation of the precursor of PhoE protein across Escherichia coli membrane vesicles requires both ATP and the protonmotive force. Biochim Biophys Acta. 1987 Jun 12;900(1):63–72. doi: 10.1016/0005-2736(87)90278-1. [DOI] [PubMed] [Google Scholar]
  15. Driessen A. J. Precursor protein translocation by the Escherichia coli translocase is directed by the protonmotive force. EMBO J. 1992 Mar;11(3):847–853. doi: 10.1002/j.1460-2075.1992.tb05122.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Elferink M. G., Hellingwerf K. J., Konings W. N. The role of the proton motive force and electron flow in solute transport in Escherichia coli. Eur J Biochem. 1985 Nov 15;153(1):161–165. doi: 10.1111/j.1432-1033.1985.tb09282.x. [DOI] [PubMed] [Google Scholar]
  17. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. doi: 10.1016/0378-1119(86)90358-6. [DOI] [PubMed] [Google Scholar]
  18. Gardel C., Johnson K., Jacq A., Beckwith J. The secD locus of E.coli codes for two membrane proteins required for protein export. EMBO J. 1990 Oct;9(10):3209–3216. doi: 10.1002/j.1460-2075.1990.tb07519.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Geller B. L. Energy requirements for protein translocation across the Escherichia coli inner membrane. Mol Microbiol. 1991 Sep;5(9):2093–2098. doi: 10.1111/j.1365-2958.1991.tb02138.x. [DOI] [PubMed] [Google Scholar]
  20. Geller B. L., Green H. M. Translocation of pro-OmpA across inner membrane vesicles of Escherichia coli occurs in two consecutive energetically distinct steps. J Biol Chem. 1989 Oct 5;264(28):16465–16469. [PubMed] [Google Scholar]
  21. Gutierrez C., Devedjian J. C. A plasmid facilitating in vitro construction of phoA gene fusions in Escherichia coli. Nucleic Acids Res. 1989 May 25;17(10):3999–3999. doi: 10.1093/nar/17.10.3999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Guzman L. M., Barondess J. J., Beckwith J. FtsL, an essential cytoplasmic membrane protein involved in cell division in Escherichia coli. J Bacteriol. 1992 Dec;174(23):7716–7728. [PMC free article] [PubMed] [Google Scholar]
  23. Hanada M., Nishiyama K. I., Mizushima S., Tokuda H. Reconstitution of an efficient protein translocation machinery comprising SecA and the three membrane proteins, SecY, SecE, and SecG (p12). J Biol Chem. 1994 Sep 23;269(38):23625–23631. [PubMed] [Google Scholar]
  24. Hartl F. U., Lecker S., Schiebel E., Hendrick J. P., Wickner W. The binding cascade of SecB to SecA to SecY/E mediates preprotein targeting to the E. coli plasma membrane. Cell. 1990 Oct 19;63(2):269–279. doi: 10.1016/0092-8674(90)90160-g. [DOI] [PubMed] [Google Scholar]
  25. Higgins D. E., DiRita V. J. Transcriptional control of toxT, a regulatory gene in the ToxR regulon of Vibrio cholerae. Mol Microbiol. 1994 Oct;14(1):17–29. doi: 10.1111/j.1365-2958.1994.tb01263.x. [DOI] [PubMed] [Google Scholar]
  26. Hoekstra W. P., Bergmans J. E., Zuidweg E. M. Role of recBC nuclease in Escherichia coli transformation. J Bacteriol. 1980 Aug;143(2):1031–1032. doi: 10.1128/jb.143.2.1031-1032.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ito K., Bassford P. J., Jr, Beckwith J. Protein localization in E. coli: is there a common step in the secretion of periplasmic and outer-membrane proteins? Cell. 1981 Jun;24(3):707–717. doi: 10.1016/0092-8674(81)90097-0. [DOI] [PubMed] [Google Scholar]
  28. Kawasaki S., Mizushima S., Tokuda H. Membrane vesicles containing overproduced SecY and SecE exhibit high translocation ATPase activity and countermovement of protons in a SecA- and presecretory protein-dependent manner. J Biol Chem. 1993 Apr 15;268(11):8193–8198. [PubMed] [Google Scholar]
  29. Kleerebezem M., Tommassen J. Expression of the pspA gene stimulates efficient protein export in Escherichia coli. Mol Microbiol. 1993 Mar;7(6):947–956. doi: 10.1111/j.1365-2958.1993.tb01186.x. [DOI] [PubMed] [Google Scholar]
  30. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  31. Kumamoto C. A. Escherichia coli SecB protein associates with exported protein precursors in vivo. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5320–5324. doi: 10.1073/pnas.86.14.5320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lill R., Cunningham K., Brundage L. A., Ito K., Oliver D., Wickner W. SecA protein hydrolyzes ATP and is an essential component of the protein translocation ATPase of Escherichia coli. EMBO J. 1989 Mar;8(3):961–966. doi: 10.1002/j.1460-2075.1989.tb03458.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lolkema J. S., Abbing A., Hellingwerf K. J., Konings W. N. The transmembrane electrical potential in Rhodopseudomonas sphaeroides determined from the distribution of tetraphenylphosphonium after correction for its binding to cell components. Eur J Biochem. 1983 Feb 1;130(2):287–292. doi: 10.1111/j.1432-1033.1983.tb07149.x. [DOI] [PubMed] [Google Scholar]
  34. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  35. Matsuyama S., Fujita Y., Mizushima S. SecD is involved in the release of translocated secretory proteins from the cytoplasmic membrane of Escherichia coli. EMBO J. 1993 Jan;12(1):265–270. doi: 10.1002/j.1460-2075.1993.tb05652.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Milstein C., Brownlee G. G., Harrison T. M., Mathews M. B. A possible precursor of immunoglobulin light chains. Nat New Biol. 1972 Sep 27;239(91):117–120. doi: 10.1038/newbio239117a0. [DOI] [PubMed] [Google Scholar]
  37. Schatz P. J., Beckwith J. Genetic analysis of protein export in Escherichia coli. Annu Rev Genet. 1990;24:215–248. doi: 10.1146/annurev.ge.24.120190.001243. [DOI] [PubMed] [Google Scholar]
  38. Schiebel E., Driessen A. J., Hartl F. U., Wickner W. Delta mu H+ and ATP function at different steps of the catalytic cycle of preprotein translocase. Cell. 1991 Mar 8;64(5):927–939. doi: 10.1016/0092-8674(91)90317-r. [DOI] [PubMed] [Google Scholar]
  39. Schiebel E., Wickner W. Preprotein translocation creates a halide anion permeability in the Escherichia coli plasma membrane. J Biol Chem. 1992 Apr 15;267(11):7505–7510. [PubMed] [Google Scholar]
  40. Schnaitman C. A. Outer membrane proteins of Escherichia coli. 3. Evidence that the major protein of Escherichia coli O111 outer membrane consists of four distinct polypeptide species. J Bacteriol. 1974 May;118(2):442–453. doi: 10.1128/jb.118.2.442-453.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Simon S. M., Blobel G. Signal peptides open protein-conducting channels in E. coli. Cell. 1992 May 15;69(4):677–684. doi: 10.1016/0092-8674(92)90231-z. [DOI] [PubMed] [Google Scholar]
  42. Tani K., Tokuda H., Mizushima S. Translocation of ProOmpA possessing an intramolecular disulfide bridge into membrane vesicles of Escherichia coli. Effect of membrane energization. J Biol Chem. 1990 Oct 5;265(28):17341–17347. [PubMed] [Google Scholar]
  43. Tommassen J., Lugtenberg B. Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase. J Bacteriol. 1980 Jul;143(1):151–157. doi: 10.1128/jb.143.1.151-157.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tommassen J., de Kroon T. Subcellular localization of a PhoE-LacZ fusion protein in E. coli by protease accessibility experiments reveals an inner-membrane-spanning form of the protein. FEBS Lett. 1987 Sep 14;221(2):226–230. doi: 10.1016/0014-5793(87)80930-4. [DOI] [PubMed] [Google Scholar]
  45. Tommassen J., van Tol H., Lugtenberg B. The ultimate localization of an outer membrane protein of Escherichia coli K-12 is not determined by the signal sequence. EMBO J. 1983;2(8):1275–1279. doi: 10.1002/j.1460-2075.1983.tb01581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  47. Weiner L., Brissette J. L., Model P. Stress-induced expression of the Escherichia coli phage shock protein operon is dependent on sigma 54 and modulated by positive and negative feedback mechanisms. Genes Dev. 1991 Oct;5(10):1912–1923. doi: 10.1101/gad.5.10.1912. [DOI] [PubMed] [Google Scholar]
  48. Weiner L., Model P. Role of an Escherichia coli stress-response operon in stationary-phase survival. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2191–2195. doi: 10.1073/pnas.91.6.2191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yamada H., Tokuda H., Mizushima S. Proton motive force-dependent and -independent protein translocation revealed by an efficient in vitro assay system of Escherichia coli. J Biol Chem. 1989 Jan 25;264(3):1723–1728. [PubMed] [Google Scholar]
  50. de Cock H., Tommassen J. SecB-binding does not maintain the translocation-competent state of prePhoE. Mol Microbiol. 1992 Mar;6(5):599–604. doi: 10.1111/j.1365-2958.1992.tb01506.x. [DOI] [PubMed] [Google Scholar]
  51. von Heijne G. Membrane protein structure prediction. Hydrophobicity analysis and the positive-inside rule. J Mol Biol. 1992 May 20;225(2):487–494. doi: 10.1016/0022-2836(92)90934-c. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES