1887

Abstract

The subcellular localization of membrane proteins in was examined by using fluorescent protein fusions. ATP synthase and succinate dehydrogenase were found to localize within discrete domains on the membrane rather than being homogeneously distributed around the cell periphery as expected. Dual labelling of cells indicated partial colocalization of ATP synthase and succinate dehydrogenase. Further analysis using an ectopically expressed phage protein gave the same localization patterns as ATP synthase and succinate dehydrogenase, implying that membrane proteins are restricted to domains within the membrane. 3D reconstruction of images of the localization of ATP synthase showed that domains were not regular and there was no bias for localization to cell poles or any other positions. Further analysis revealed that this localization was highly dynamic, but random, implying that integral membrane proteins are free to diffuse two-dimensionally around the cytoplasmic membrane.

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2004-09-01
2019-11-20
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References

  1. Anagnostopoulos, C. & Spizizen, J. ( 1961; ). Requirements for transformation in Bacillus subtilis. J Bacteriol 81, 741–746.
    [Google Scholar]
  2. Brandon, L. D., Goehring, N., Janakirman, A., Yan, A. W., Wu, T., Beckwith, J. & Goldberg, M. B. ( 2003; ). IcsA, a polarly localized autotransporter with an atypical signal peptide, uses the Sec apparatus for secretion, although the Sec apparatus is circumferentially distributed. Mol Microbiol 50, 45–60.[CrossRef]
    [Google Scholar]
  3. Carballido-López, R. & Errington, J. ( 2003; ). The bacterial cytoskeleton: in vivo dynamics of the actin-like protein Mbl of Bacillus subtilis. Dev Cell 4, 19–28.[CrossRef]
    [Google Scholar]
  4. de Mendoza, D., Schujman, G. E. & Aguilar, P. S. ( 2002; ). Biosynthesis and function of membrane lipids. In Bacillus subtilis and its Closest Relatives: from Genes to Cells, pp. 43–56. Edited by A. L. Sonenshine, J. A. Hoch & R. Losick. Washington, DC: American Society for Microbiology Press.
  5. Edidin, M. ( 2003; ). The state of lipid rafts: from model membranes to cells. Annu Rev Biophys Biomol Struct 32, 257–283.[CrossRef]
    [Google Scholar]
  6. Feucht, A. & Lewis, P. J. ( 2001; ). Improved plasmid vectors for the production of multiple fluorescent protein fusions in Bacillus subtilis. Gene 264, 289–297.[CrossRef]
    [Google Scholar]
  7. Fishov, I. & Woldringh, C. ( 1999; ). Visualization of membrane domains in Escherichia coli. Mol Microbiol 32, 1166–1172.[CrossRef]
    [Google Scholar]
  8. Glaser, P., Sharpe, M. E., Raether, B., Perego, M., Ohlsen, K. & Errington, J. ( 1997; ). Dynamic, mitotic-like behaviour of a bacterial protein required for accurate chromosome partitioning. Genes Dev 11, 1160–1168.[CrossRef]
    [Google Scholar]
  9. Jenkinson, H. ( 1983; ). Altered arrangement of proteins in the spore coat of a germination mutant of Bacillus subtilis. J Gen Microbiol 129, 1945–1958.
    [Google Scholar]
  10. Jones, L. J. F., Carballido-López, R. & Errington, J. ( 2001; ). Control of cell shape in bacteria: helical, actin-like filaments in Bacillus subtilis. Cell 104, 913–922.[CrossRef]
    [Google Scholar]
  11. Kadner, R. J. ( 1996; ). Cytoplasmic membrane. In Escherichia coli and Salmonella: Cellular and Molecular Biology 2nd edn, pp 58–87. Edited by F. C. Neidhardt and others. Washington DC: American Society for Microbiology.
  12. Kawai, F., Shoda, M., Harashima, R., Sadaie, Y., Hara, H. & Matsumoto, K. ( 2004; ). Cardiolipin domains in Bacillus subtilis Marburg membranes. J Bacteriol 186, 1475–1483.[CrossRef]
    [Google Scholar]
  13. Kruse, T., Møller-Jensen, J., Løbner-Oelsen, A. & Gerdes, K. ( 2003; ). Dysfunctional MreB inhibits chromosome segregation in Escherichia coli. EMBO J 22, 5283–5292.[CrossRef]
    [Google Scholar]
  14. Lewis, P. J. & Marston, A. L. ( 1999; ). GFP vectors for controlled expression and dual labelling of protein fusions in Bacillus subtilis. Gene 227, 101–109.[CrossRef]
    [Google Scholar]
  15. Lewis, P. J., Thaker, S. D. & Errington, J. ( 2000; ). Compartmentalization of transcription and translation in Bacillus subtilis. EMBO J 19, 710–718.[CrossRef]
    [Google Scholar]
  16. Lopian, L., Nussbaum-Schochat, A., O'Day-Kerstein, K., Wright, A. & Amster-Choder, O. ( 2003; ). The BglF sensor recruits the BglG transcription regulator to the membrane and releases it on stimulation. Proc Natl Acad Sci U S A 100, 7099–7104.[CrossRef]
    [Google Scholar]
  17. Maddock, J. R. & Shapiro, L. ( 1993; ). Polar location of the chemoreceptor complex in the Escherichia coli cell. Science 19, 1717–1723.
    [Google Scholar]
  18. Meijer, W. J. J., Serna-Rico, A. & Salas, M. ( 2001; ). Characterisation of the bacteriophage π29-encoded protein p16.7: a membrane protein involved in phage DNA replication. Mol Microbiol 39, 731–746.[CrossRef]
    [Google Scholar]
  19. Mileykovskaya, E. & Dowhan, W. ( 2000; ). Visualization of phospholipid domains in Escherichia coli by using the cardiolipin-specific fluorescent dye 10-N-nonyl acridine orange. J Bacteriol 182, 1172–1175.[CrossRef]
    [Google Scholar]
  20. Minaschek, G., Groschel-Stewart, U., Blum, S. & Bereiter-Hahn, J. ( 1992; ). Microcompartmentation of glycolytic enzymes in cultured cells. Eur J Cell Biol 58, 418–428.
    [Google Scholar]
  21. Pogliano, J., Osborne, N., Sharp, M. D., Abanes-De Mello, A., Perez, A., Sun, Y. L. & Pogliano, K. ( 1999; ). A vital stain for studying membrane dynamics in bacteria: a novel mechanism controlling septation during Bacillus subtilis sporulation. Mol Microbiol 31, 1149–1159.[CrossRef]
    [Google Scholar]
  22. Santana, M., Ionescu, M. S., Vertes, A., Longin, R., Kunst, F., Danchin, A. & Glaser, P. ( 1994; ). Bacillus subtilis F0F1 ATPase: DNA sequence of the atp operon and characterization of atp mutants. J Bacteriol 176, 6802–6811.
    [Google Scholar]
  23. Schägger, H. ( 2002; ). Respiratory chain supercomplexes of mitochondria and bacteria. Biochim Biophys Acta 155, 154–159.
    [Google Scholar]
  24. Scheffers, D.-J., Jones, L. J. F. & Errington, J. ( 2004; ). Several distinct localization patterns for penicillin-binding proteins in Bacillus subtilis. Mol Microbiol 51, 749–764.
    [Google Scholar]
  25. Sharpe, M. E., Hauser, P. M., Sharpe, R. G. & Errington, J. ( 1998; ). Bacillus subtilis cell cycle as studied by microscopy: constancy of cell length at initiation of DNA replication and evidence for active nucleoid partitioning. J Bacteriol 180, 547–555.
    [Google Scholar]
  26. Vanounou, S., Parola, A. H. & Fishov, I. ( 2003; ). Phosphatidylethanolamine and phosphatidylglycerol are segregated into different domains in bacterial membrane. A study with pyrene-labelled phospholipids. Mol Microbiol 49, 1067–1079.[CrossRef]
    [Google Scholar]
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Movie of 3D reconstruction of AtpA-GFP fusion shown in Fig. 4 of the main text. Movie of 30 min time-lapse monitoring of AtpA-GFP movement described in the main text and shown in Fig. 5 of the main text. Each frame represents a 1 min time gap. Analysis of ATP synthase movement over a short time period. Panels A-C show separate frames from a time-lapse experiment taken 5 s apart. Panel D shows an overlay of the three frames. Panel E shows a linescan through the cell illustrated with the arrow in panel D, with the same colour scheme for 0 s (red), 5 s (green) and 10 s (blue) in all panels. The majority of the peaks overlap at each time point, indicating that there was little movement of ATP synthase over a 10 s period.

MOVIE

Movie of 3D reconstruction of AtpA-GFP fusion shown in Fig. 4 of the main text. Movie of 30 min time-lapse monitoring of AtpA-GFP movement described in the main text and shown in Fig. 5 of the main text. Each frame represents a 1 min time gap. Analysis of ATP synthase movement over a short time period. Panels A-C show separate frames from a time-lapse experiment taken 5 s apart. Panel D shows an overlay of the three frames. Panel E shows a linescan through the cell illustrated with the arrow in panel D, with the same colour scheme for 0 s (red), 5 s (green) and 10 s (blue) in all panels. The majority of the peaks overlap at each time point, indicating that there was little movement of ATP synthase over a 10 s period.

MOVIE

vol. , part 9, pp. 2815 - 2824

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