1887

Abstract

The spore is surrounded by a 220 μm thick trilaminar coat that consists of inner and outer electron-dense layers surrounding a central region of cellulose microfibrils. In previous studies, a mutant strain (TL56) lacking three proteins associated with the outer layer exhibited increased permeability to macromolecular tracers, suggesting that this layer contributes to the coat permeability barrier. Electron microscopy now shows that the outer layer is incomplete in the coats of this mutant and consists of a residual regular array of punctate electron densities. The outer layer is also incomplete in a mutant lacking a cellulose-binding protein associated with the inner layer, and these coats are deficient in an outer-layer protein and another coat protein. To examine the mechanism by which this inner-layer protein, SP85, contributes to outer-layer formation, various domain fragments were overexpressed in forming spores. Most of these exert dominant negative effects similar to the deletion of outer-layer proteins, but one construct, consisting of a fusion of the N-terminal and Cys-rich C1 domain, induces a dense mat of novel filaments at the surface of the outer layer. Biochemical studies show that the C1 domain binds cellulose, and a combination of site-directed mutations that inhibits its cellulose-binding activity suppresses outer-layer filament induction. The results suggest that, in addition to a previously described early role in regulating cellulose synthesis, SP85 subsequently contributes a cross-bridging function between cellulose and other coat proteins to organize previously unrecognized structural elements in the outer layer of the coat.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.25984-0
2003-02-01
2019-10-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/2/mic149305.html?itemId=/content/journal/micro/10.1099/mic.0.25984-0&mimeType=html&fmt=ahah

References

  1. Aparicio, J. G., Erdos, G. W. & West, C. M. ( 1990; ). Spore coat is altered in modB glycosylation mutants of Dictyostelium discoideum. J Cell Biochem 42, 255–266.[CrossRef]
    [Google Scholar]
  2. Bayer, E. A., Shimon, L. J., Shoham, Y. & Lamed, R. ( 1998; ). Cellulosomes – structure and ultrastructure. J Struct Biol 124, 221–234.[CrossRef]
    [Google Scholar]
  3. Blanton, R. L., Fuller, D., Iranfar, N., Grimson, M. J. & Loomis, W. F. ( 2000; ). The cellulose synthase gene of Dictyostelium. Proc Natl Acad Sci U S A 97, 2391–2396.[CrossRef]
    [Google Scholar]
  4. Boulianne, R. P., Liu, Y., Aebi, M., Lu, B. C. & Kues, U. ( 2000; ). Fruiting body development in Coprinus cinerus: regulated expression of two galectins secreted by a non-classical pathway. Microbiology 146, 1841–1853.
    [Google Scholar]
  5. Brett, C. T. ( 2000; ). Cellulose microfibrils in plants: biosynthesis, deposition, and integration into the cell wall. Int Rev Cyt 199, 161–199.
    [Google Scholar]
  6. Delmer, D. P. ( 1999; ). Cellulose biosynthesis: exciting times for a difficult field of study. Annu Rev Plant Physiol Plant Mol Biol 50, 245–276.[CrossRef]
    [Google Scholar]
  7. Erdos, G. W. & West, C. M. ( 1989; ). Formation and organization of the spore coat of Dictyostelium discoideum. Exp Mycol 13, 169–182.[CrossRef]
    [Google Scholar]
  8. Fosnaugh, K. L., Fuller, D. & Loomis, W. F. ( 1995; ). Structural roles of the spore coat proteins in Dictyostelium discoideum. Dev Biol 166, 823–825.
    [Google Scholar]
  9. Frisardi, M., Ghosh, S. K., Field, J., VanDellen, K., Rogers, R., Robbins, P. & Samuelson, J. ( 2000; ). The most abundant glycoprotein of amebic cyst walls (Jacob) is a lectin with five Cys-rich, chitin-binding domains. Infect Immun 68, 4217–4224.[CrossRef]
    [Google Scholar]
  10. Gonzalez-Yanes, B., Mandell, R. B., Girard, M., Henry, S., Aparicio, O., Gritzali, M., Brown, R. D., Erdos, G. W. & West, C. M. ( 1989; ). The spore coat of a fucosylation mutant in Dictyostelium discoideum. Dev Biol 133, 576–587.[CrossRef]
    [Google Scholar]
  11. Handford, P. A., Downing, A. K., Reinhardt, D. P. & Sakai, L. Y. ( 2000; ). Fibrillin: from domain structure to supramolecular assembly. Matrix Biol 19, 457–470.[CrossRef]
    [Google Scholar]
  12. Harb, O. S., Gao, L.-Y. & Abu Kwaik, Y. ( 2000; ). From protozoa to mammalian cells: a new paradigm in the life cycle of intracellular bacterial pathogens. Environ Microbiol 2, 251–265.[CrossRef]
    [Google Scholar]
  13. Kelly, M. A., Chellgren, B. W., Rucker, A. L., Troutman, J. M., Fried, M. G., Miller, A.-F. & Creamer, T. P. ( 2001; ). Host–guest study of left-handed polyproline II helix formation. Biochemistry 40, 14376–14383.[CrossRef]
    [Google Scholar]
  14. Loomis, W. F. ( 1971; ). Sensitivity of Dictyostelium discoideum to nucleic acid analogues. Exp Cell Res 64, 484–486.[CrossRef]
    [Google Scholar]
  15. Maeda, M. ( 1992; ). Efficient induction of sporulation of Dictyostelium prespore cells by 8-bromocyclic AMP under both submerged- and shaken-culture conditions and involvement of protein kinase(s) in its action. Dev Growth Differ 34, 263–275.[CrossRef]
    [Google Scholar]
  16. Mateos, F. V., Rickauer, M. & Esquerre-Tugaye, M.-T. ( 1997; ). Cloning and characterization of a cDNA encoding an elicitor of Phytophthora parasitica var. nicotianae that shows cellulose-binding and lectin-like activities. Mol Plant Microbe Interact 10, 1045–1053.[CrossRef]
    [Google Scholar]
  17. McDowell, E. M. & Trump, B. F. ( 1976; ). Histologic fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 100, 405–414.
    [Google Scholar]
  18. McGuire, V. & Alexander, S. ( 1996; ). PsB multiprotein complex of Dictyostelium discoideum: demonstration of cellulose binding activity and order of protein subunit assembly. J Biol Chem 271, 14596–14603.[CrossRef]
    [Google Scholar]
  19. Nakao, H., Yamamoto, A., Takeuchi, I. & Tasaka, M. ( 1994; ). Dictyostelium prespore-specific gene (DP87) encodes a sorus matrix protein. J Cell Sci 107, 397–403.
    [Google Scholar]
  20. Orlowski, M. & Loomis, W. F. ( 1979; ). Plasma membrane proteins of Dictyostelium: the spore coat proteins. Dev Biol 71, 297–307.[CrossRef]
    [Google Scholar]
  21. Pierini, L. M. & Doering, T. L. ( 2001; ). Spatial and temporal sequence of capsule construction in Cryptococcus neoformans. Mol Microbiol 41, 105–115.[CrossRef]
    [Google Scholar]
  22. Sassi, S., Sweetinburgh, M., Erogul, J., Zhang, P., Teng-umnuay, P. & West, C. M. ( 2001; ). Analysis of Skp1 glycosylation and nuclear enrichment in Dictyostelium. Glycobiology 11, 283–295.[CrossRef]
    [Google Scholar]
  23. Simrell, C. R. & Klein, P. A. ( 1979; ). Antibody responses of tumor-bearing mice to their own tumors captured and perpetuated as hybridomas. J Immunol 123, 2386–2394.
    [Google Scholar]
  24. Srinivasan, S., Griffiths, K. R., McGuire, V., Champion, A., Williams, K. L. & Alexander, S. ( 2000; ). The cellulose-binding activity of the PsB multiprotein complex is required for proper assembly of the spore coat and spore viability in Dictyostelium discoideum. Microbiology 146, 1829–1839.
    [Google Scholar]
  25. Tani, P. H., Loftus, J. C. & Bowditch, R. D. ( 2002; ). In vitro selection of fibronectin gain-of-function mutations. Biochem J 365, 287–294.[CrossRef]
    [Google Scholar]
  26. Wang, W. & Malcolm, B. A. ( 1999; ). Two-stage PCR protocol allowing introduction of multiple mutations, deletions and insertions using QuikChange site-directed mutagenesis. Biotechniques 26, 680–682.
    [Google Scholar]
  27. Wang, Y., Slade, M. B., Gooley, A. A., Atwell, B. J. & Williams, K. L. ( 2001; ). Cellulose-binding modules from extracellular matrix proteins of Dictyostelium discoideum stalk and sheath. Eur J Biochem 268, 4334–4345.[CrossRef]
    [Google Scholar]
  28. West, C. M. ( 2002; ). Comparative analysis of spore coat formation, structure and function in Dictyostelium. Int Rev Cyt 222, 237–293.
    [Google Scholar]
  29. West, C. M. & Erdos, G. W. ( 1988; ). The expression of glycoproteins in the extracellular matrix of the cellular slime mold Dictyostelium discoideum. Cell Differ 23, 1–16.[CrossRef]
    [Google Scholar]
  30. West, C. M., Mao, J., van der Wel, H., Erdos, G. W. & Zhang, Y. ( 1996; ). SP75 is encoded by the DP87 gene and belongs to a family of modular Dictyostelium discoideum outer layer spore coat proteins. Microbiology 142, 2227–2243.[CrossRef]
    [Google Scholar]
  31. West, C. M., Zhang, P., McGlynn, A. C. & Kaplan, L. ( 2002; ). Outside–in signaling of cellulose synthesis by a spore coat protein in Dictyostelium. Euk Cell 1, 281–292.[CrossRef]
    [Google Scholar]
  32. Woessner, J. P. & Goodenough, U. W. ( 1994; ). Volvocine cell walls and their constituent glycoproteins: an evolutionary perspective. Protoplasma 181, 245–258.[CrossRef]
    [Google Scholar]
  33. Zhang, P., McGlynn, A. C., Loomis, W. F., Blanton, R. L. & West, C. M. ( 2001; ). Spore coat formation and timely sporulation depend on cellulose in Dictyostelium. Differentation 67, 72–79.[CrossRef]
    [Google Scholar]
  34. Zhang, Y., Brown, R. D. & West, C. M. ( 1998; ). Two proteins of the Dictyostelium spore coat bind to cellulose in vitro. Biochemistry 37, 10766–10779.[CrossRef]
    [Google Scholar]
  35. Zhang, Y., Zhang, P. & West, C. M. ( 1999; ). A linking function for the cellulose-binding protein SP85 in the spore coat of Dictyostelium discoideum. J Cell Sci 112, 4367–4377.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.25984-0
Loading
/content/journal/micro/10.1099/mic.0.25984-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error