Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Aug;179(16):5094–5103. doi: 10.1128/jb.179.16.5094-5103.1997

Domain-swapping analysis of FtsI, FtsL, and FtsQ, bitopic membrane proteins essential for cell division in Escherichia coli.

L M Guzman 1, D S Weiss 1, J Beckwith 1
PMCID: PMC179367  PMID: 9260951

Abstract

FtsI, FtsL, and FtsQ are three membrane proteins required for assembly of the division septum in the bacterium Escherichia coli. Cells lacking any of these three proteins form long, aseptate filaments that eventually lyse. FtsI, FtsL, and FtsQ are not homologous but have similar overall structures: a small cytoplasmic domain, a single membrane-spanning segment (MSS), and a large periplasmic domain that probably encodes the primary functional activities of these proteins. The periplasmic domain of FtsI catalyzes transpeptidation and is involved in the synthesis of septal peptidoglycan. The precise functions of FtsL and FtsQ are not known. To ask whether the cytoplasmic domain and MSS of each protein serve only as a membrane anchor or have instead a more sophisticated function, we have used molecular genetic techniques to swap these domains among the three Fts proteins and one membrane protein not involved in cell division, MalF. In the cases of FtsI and FtsL, replacement of the cytoplasmic domain and/or MSS resulted in the loss of the ability to support cell division. For FtsQ, MSS swaps supported cell division but cytoplasmic domain swaps did not. We discuss several potential interpretations of these results, including that the essential domains of FtsI, FtsL, and FtsQ have a role in regulating the localization and/or activity of these proteins to ensure that septum formation occurs at the right place in the cell and at the right time during the division cycle.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Barondess J. J., Carson M., Guzman Verduzco L. M., Beckwith J. Alkaline phosphatase fusions in the study of cell division genes. Res Microbiol. 1991 Feb-Apr;142(2-3):295–299. doi: 10.1016/0923-2508(91)90044-b. [DOI] [PubMed] [Google Scholar]
  2. Bi E. F., Lutkenhaus J. FtsZ ring structure associated with division in Escherichia coli. Nature. 1991 Nov 14;354(6349):161–164. doi: 10.1038/354161a0. [DOI] [PubMed] [Google Scholar]
  3. Bonifacino J. S., Cosson P., Shah N., Klausner R. D. Role of potentially charged transmembrane residues in targeting proteins for retention and degradation within the endoplasmic reticulum. EMBO J. 1991 Oct;10(10):2783–2793. doi: 10.1002/j.1460-2075.1991.tb07827.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowler L. D., Spratt B. G. Membrane topology of penicillin-binding protein 3 of Escherichia coli. Mol Microbiol. 1989 Sep;3(9):1277–1286. doi: 10.1111/j.1365-2958.1989.tb00278.x. [DOI] [PubMed] [Google Scholar]
  5. Boyd D., Beckwith J. Positively charged amino acid residues can act as topogenic determinants in membrane proteins. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9446–9450. doi: 10.1073/pnas.86.23.9446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boyd D., Manoil C., Beckwith J. Determinants of membrane protein topology. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8525–8529. doi: 10.1073/pnas.84.23.8525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carson M. J., Barondess J., Beckwith J. The FtsQ protein of Escherichia coli: membrane topology, abundance, and cell division phenotypes due to overproduction and insertion mutations. J Bacteriol. 1991 Apr;173(7):2187–2195. doi: 10.1128/jb.173.7.2187-2195.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cosson P., Lankford S. P., Bonifacino J. S., Klausner R. D. Membrane protein association by potential intramembrane charge pairs. Nature. 1991 May 30;351(6325):414–416. doi: 10.1038/351414a0. [DOI] [PubMed] [Google Scholar]
  9. Dai K., Lutkenhaus J. The proper ratio of FtsZ to FtsA is required for cell division to occur in Escherichia coli. J Bacteriol. 1992 Oct;174(19):6145–6151. doi: 10.1128/jb.174.19.6145-6151.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dai K., Xu Y., Lutkenhaus J. Topological characterization of the essential Escherichia coli cell division protein FtsN. J Bacteriol. 1996 Mar;178(5):1328–1334. doi: 10.1128/jb.178.5.1328-1334.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dougherty T. J., Kennedy K., Kessler R. E., Pucci M. J. Direct quantitation of the number of individual penicillin-binding proteins per cell in Escherichia coli. J Bacteriol. 1996 Nov;178(21):6110–6115. doi: 10.1128/jb.178.21.6110-6115.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ehrmann M., Beckwith J. Proper insertion of a complex membrane protein in the absence of its amino-terminal export signal. J Biol Chem. 1991 Sep 5;266(25):16530–16533. [PubMed] [Google Scholar]
  13. Fraipont C., Adam M., Nguyen-Distèche M., Keck W., Van Beeumen J., Ayala J. A., Granier B., Hara H., Ghuysen J. M. Engineering and overexpression of periplasmic forms of the penicillin-binding protein 3 of Escherichia coli. Biochem J. 1994 Feb 15;298(Pt 1):189–195. doi: 10.1042/bj2980189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Froshauer S., Beckwith J. The nucleotide sequence of the gene for malF protein, an inner membrane component of the maltose transport system of Escherichia coli. Repeated DNA sequences are found in the malE-malF intercistronic region. J Biol Chem. 1984 Sep 10;259(17):10896–10903. [PubMed] [Google Scholar]
  15. Ghuysen J. M. Molecular structures of penicillin-binding proteins and beta-lactamases. Trends Microbiol. 1994 Oct;2(10):372–380. doi: 10.1016/0966-842x(94)90614-9. [DOI] [PubMed] [Google Scholar]
  16. Goffin C., Fraipont C., Ayala J., Terrak M., Nguyen-Distèche M., Ghuysen J. M. The non-penicillin-binding module of the tripartite penicillin-binding protein 3 of Escherichia coli is required for folding and/or stability of the penicillin-binding module and the membrane-anchoring module confers cell septation activity on the folded structure. J Bacteriol. 1996 Sep;178(18):5402–5409. doi: 10.1128/jb.178.18.5402-5409.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Guarnieri F. G., Arterburn L. M., Penno M. B., Cha Y., August J. T. The motif Tyr-X-X-hydrophobic residue mediates lysosomal membrane targeting of lysosome-associated membrane protein 1. J Biol Chem. 1993 Jan 25;268(3):1941–1946. [PubMed] [Google Scholar]
  18. 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]
  19. Guzman L. M., Belin D., Carson M. J., Beckwith J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol. 1995 Jul;177(14):4121–4130. doi: 10.1128/jb.177.14.4121-4130.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Heijne G. The distribution of positively charged residues in bacterial inner membrane proteins correlates with the trans-membrane topology. EMBO J. 1986 Nov;5(11):3021–3027. doi: 10.1002/j.1460-2075.1986.tb04601.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Henderson R., Baldwin J. M., Ceska T. A., Zemlin F., Beckmann E., Downing K. H. Model for the structure of bacteriorhodopsin based on high-resolution electron cryo-microscopy. J Mol Biol. 1990 Jun 20;213(4):899–929. doi: 10.1016/S0022-2836(05)80271-2. [DOI] [PubMed] [Google Scholar]
  22. Hofnung M. Divergent operons and the genetic structure of the maltose B region in Escherichia coli K12. Genetics. 1974 Feb;76(2):169–184. doi: 10.1093/genetics/76.2.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ishino F., Matsuhashi M. Peptidoglycan synthetic enzyme activities of highly purified penicillin-binding protein 3 in Escherichia coli: a septum-forming reaction sequence. Biochem Biophys Res Commun. 1981 Aug 14;101(3):905–911. doi: 10.1016/0006-291x(81)91835-0. [DOI] [PubMed] [Google Scholar]
  24. Lakaye B., Damblon C., Jamin M., Galleni M., Lepage S., Joris B., Marchand-Brynaert J., Frydrych C., Frere J. M. Synthesis, purification and kinetic properties of fluorescein-labelled penicillins. Biochem J. 1994 May 15;300(Pt 1):141–145. doi: 10.1042/bj3000141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lemmon M. A., Flanagan J. M., Hunt J. F., Adair B. D., Bormann B. J., Dempsey C. E., Engelman D. M. Glycophorin A dimerization is driven by specific interactions between transmembrane alpha-helices. J Biol Chem. 1992 Apr 15;267(11):7683–7689. [PubMed] [Google Scholar]
  26. McGovern K., Ehrmann M., Beckwith J. Decoding signals for membrane protein assembly using alkaline phosphatase fusions. EMBO J. 1991 Oct;10(10):2773–2782. doi: 10.1002/j.1460-2075.1991.tb07826.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mukherjee A., Donachie W. D. Differential translation of cell division proteins. J Bacteriol. 1990 Oct;172(10):6106–6111. doi: 10.1128/jb.172.10.6106-6111.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Murphy C. K., Beckwith J. Residues essential for the function of SecE, a membrane component of the Escherichia coli secretion apparatus, are located in a conserved cytoplasmic region. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2557–2561. doi: 10.1073/pnas.91.7.2557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nagasawa H., Sakagami Y., Suzuki A., Suzuki H., Hara H., Hirota Y. Determination of the cleavage site involved in C-terminal processing of penicillin-binding protein 3 of Escherichia coli. J Bacteriol. 1989 Nov;171(11):5890–5893. doi: 10.1128/jb.171.11.5890-5893.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nilsson I., von Heijne G. Fine-tuning the topology of a polytopic membrane protein: role of positively and negatively charged amino acids. Cell. 1990 Sep 21;62(6):1135–1141. doi: 10.1016/0092-8674(90)90390-z. [DOI] [PubMed] [Google Scholar]
  31. Nilsson T., Lucocq J. M., Mackay D., Warren G. The membrane spanning domain of beta-1,4-galactosyltransferase specifies trans Golgi localization. EMBO J. 1991 Dec;10(12):3567–3575. doi: 10.1002/j.1460-2075.1991.tb04923.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nothwehr S. F., Roberts C. J., Stevens T. H. Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues. J Cell Biol. 1993 Jun;121(6):1197–1209. doi: 10.1083/jcb.121.6.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pelham H. R., Munro S. Sorting of membrane proteins in the secretory pathway. Cell. 1993 Nov 19;75(4):603–605. doi: 10.1016/0092-8674(93)90479-A. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Popham D. L., Setlow P. Phenotypes of Bacillus subtilis mutants lacking multiple class A high-molecular-weight penicillin-binding proteins. J Bacteriol. 1996 Apr;178(7):2079–2085. doi: 10.1128/jb.178.7.2079-2085.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pringle J. R., Adams A. E., Drubin D. G., Haarer B. K. Immunofluorescence methods for yeast. Methods Enzymol. 1991;194:565–602. doi: 10.1016/0076-6879(91)94043-c. [DOI] [PubMed] [Google Scholar]
  36. Roth M. G., Doyle C., Sambrook J., Gething M. J. Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway. J Cell Biol. 1986 Apr;102(4):1271–1283. doi: 10.1083/jcb.102.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schutze M. P., Peterson P. A., Jackson M. R. An N-terminal double-arginine motif maintains type II membrane proteins in the endoplasmic reticulum. EMBO J. 1994 Apr 1;13(7):1696–1705. doi: 10.1002/j.1460-2075.1994.tb06434.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Spratt B. G., Cromie K. D. Penicillin-binding proteins of gram-negative bacteria. Rev Infect Dis. 1988 Jul-Aug;10(4):699–711. doi: 10.1093/clinids/10.4.699. [DOI] [PubMed] [Google Scholar]
  39. Strauch K. L., Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1576–1580. doi: 10.1073/pnas.85.5.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. von Heijne G. Control of topology and mode of assembly of a polytopic membrane protein by positively charged residues. Nature. 1989 Oct 5;341(6241):456–458. doi: 10.1038/341456a0. [DOI] [PubMed] [Google Scholar]
  41. 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 Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES