Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Sep;176(17):5372–5377. doi: 10.1128/jb.176.17.5372-5377.1994

Secretion of the Serratia marcescens HasA protein by an ABC transporter.

S Létoffé 1, J M Ghigo 1, C Wandersman 1
PMCID: PMC196723  PMID: 8071214

Abstract

We previously identified a Serratia marcescens extracellular protein, HasA, able to bind heme and required for iron acquisition from heme and hemoglobin by the bacterium. This novel type of extracellular protein does not have a signal peptide and does not show sequence similarities to other proteins. HasA secretion was reconstituted in Escherichia coli, and we show here that like many proteins lacking a signal peptide, HasA has a C-terminal targeting sequence and is secreted by a specific ATP binding cassette (ABC) transporter consisting of three proteins, one inner membrane protein with a conserved ATP binding domain, called the ABC; a second inner membrane protein; and a third, outer membrane component. Since the three S. marcescens components of the HasA transporter have not yet been identified, the reconstituted HasA secretion system is a hybrid. It consists of the two S. marcescens inner membrane-specific components, HasD and HasE, associated with an outer membrane component coming from another bacterial ABC transporter, such as the E. coli TolC protein, the outer membrane component of the hemolysin transporter, or the Erwinia chrysanthemi PrtF protein, the outer membrane component of the protease transporter. This hybrid transporter was first shown to allow the secretion of the S. marcescens metalloprotease and the E. chrysanthemi metalloproteases B and C. On account of that, the two S. marcescens components HasD and HasE were previously named PrtDSM and PrtESM, respectively. However, HasA is secreted neither by the PrtD-PrtE-PrtF transporter (the genuine E. chrysanthemi protease transporter) nor by the HlyB-HlhD-TolC transporter (the hemolysin transporter). Moreover, HasA, coexpressed in the same cell, strongly inhibits the secretion of proteases B and C by their own transporter, indicating that the E. chrysanthemi transporter recognizes HasA. Since PrtF could replace TolC in the constitution of the HasA transporter, this indicates that the secretion block does not take place at the level of the outer membrane component but rather at an earlier step of interaction between HasA and the inner membrane components.

Full text

PDF
5372

Images in this article

5374Image
on p.5374
5374Image
on p.5374
5375Image
on p.5375
5375Image
on p.5375
5376Image
on p.5376

Selected References

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

  1. Akatsuka H., Kawai E., Omori K., Komatsubara S., Shibatani T., Tosa T. The lipA gene of Serratia marcescens which encodes an extracellular lipase having no N-terminal signal peptide. J Bacteriol. 1994 Apr;176(7):1949–1956. doi: 10.1128/jb.176.7.1949-1956.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angerer A., Klupp B., Braun V. Iron transport systems of Serratia marcescens. J Bacteriol. 1992 Feb;174(4):1378–1387. doi: 10.1128/jb.174.4.1378-1387.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ball T. K., Saurugger P. N., Benedik M. J. The extracellular nuclease gene of Serratia marcescens and its secretion from Escherichia coli. Gene. 1987;57(2-3):183–192. doi: 10.1016/0378-1119(87)90121-1. [DOI] [PubMed] [Google Scholar]
  4. Delepelaire P., Wandersman C. Characterization, localization and transmembrane organization of the three proteins PrtD, PrtE and PrtF necessary for protease secretion by the gram-negative bacterium Erwinia chrysanthemi. Mol Microbiol. 1991 Oct;5(10):2427–2434. doi: 10.1111/j.1365-2958.1991.tb02088.x. [DOI] [PubMed] [Google Scholar]
  5. Delepelaire P., Wandersman C. Protease secretion by Erwinia chrysanthemi. Proteases B and C are synthesized and secreted as zymogens without a signal peptide. J Biol Chem. 1989 May 25;264(15):9083–9089. [PubMed] [Google Scholar]
  6. Delepelaire P., Wandersman C. Protein secretion in gram-negative bacteria. The extracellular metalloprotease B from Erwinia chrysanthemi contains a C-terminal secretion signal analogous to that of Escherichia coli alpha-hemolysin. J Biol Chem. 1990 Oct 5;265(28):17118–17125. [PubMed] [Google Scholar]
  7. Ghigo J. M., Wandersman C. A fourth metalloprotease gene in Erwinia chrysanthemi. Res Microbiol. 1992 Nov-Dec;143(9):857–867. doi: 10.1016/0923-2508(92)90073-w. [DOI] [PubMed] [Google Scholar]
  8. Gilson L., Mahanty H. K., Kolter R. Four plasmid genes are required for colicin V synthesis, export, and immunity. J Bacteriol. 1987 Jun;169(6):2466–2470. doi: 10.1128/jb.169.6.2466-2470.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Higgins C. F. ABC transporters: from microorganisms to man. Annu Rev Cell Biol. 1992;8:67–113. doi: 10.1146/annurev.cb.08.110192.000435. [DOI] [PubMed] [Google Scholar]
  10. Hines D. A., Saurugger P. N., Ihler G. M., Benedik M. J. Genetic analysis of extracellular proteins of Serratia marcescens. J Bacteriol. 1988 Sep;170(9):4141–4146. doi: 10.1128/jb.170.9.4141-4146.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Létoffé S., Ghigo J. M., Wandersman C. Identification of two components of the Serratia marcescens metalloprotease transporter: protease SM secretion in Escherichia coli is TolC dependent. J Bacteriol. 1993 Nov;175(22):7321–7328. doi: 10.1128/jb.175.22.7321-7328.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mackman N., Nicaud J. M., Gray L., Holland I. B. Secretion of haemolysin by Escherichia coli. Curr Top Microbiol Immunol. 1986;125:159–181. doi: 10.1007/978-3-642-71251-7_10. [DOI] [PubMed] [Google Scholar]
  13. Nakahama K., Yoshimura K., Marumoto R., Kikuchi M., Lee I. S., Hase T., Matsubara H. Cloning and sequencing of Serratia protease gene. Nucleic Acids Res. 1986 Jul 25;14(14):5843–5855. doi: 10.1093/nar/14.14.5843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schiebel E., Schwarz H., Braun V. Subcellular location and unique secretion of the hemolysin of Serratia marcescens. J Biol Chem. 1989 Sep 25;264(27):16311–16320. [PubMed] [Google Scholar]
  16. Tommassen J., Filloux A., Bally M., Murgier M., Lazdunski A. Protein secretion in Pseudomonas aeruginosa. FEMS Microbiol Rev. 1992 Sep;9(1):73–90. doi: 10.1016/0378-1097(92)90336-m. [DOI] [PubMed] [Google Scholar]
  17. Wandersman C., Delepelaire P., Letoffe S., Schwartz M. Characterization of Erwinia chrysanthemi extracellular proteases: cloning and expression of the protease genes in Escherichia coli. J Bacteriol. 1987 Nov;169(11):5046–5053. doi: 10.1128/jb.169.11.5046-5053.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wandersman C., Delepelaire P. TolC, an Escherichia coli outer membrane protein required for hemolysin secretion. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4776–4780. doi: 10.1073/pnas.87.12.4776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wandersman C. Secretion across the bacterial outer membrane. Trends Genet. 1992 Sep;8(9):317–322. doi: 10.1016/0168-9525(92)90264-5. [DOI] [PubMed] [Google Scholar]
  20. White M. F., Kahn C. R. The insulin signaling system. J Biol Chem. 1994 Jan 7;269(1):1–4. [PubMed] [Google Scholar]
  21. Yanagida N., Uozumi T., Beppu T. Specific excretion of Serratia marcescens protease through the outer membrane of Escherichia coli. J Bacteriol. 1986 Jun;166(3):937–944. doi: 10.1128/jb.166.3.937-944.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES