1887

Abstract

SUMMARY

Following the removal of the rodlet layer from aerial or submerged conidia of the entomopathogenic deuteromycetous fungus , SDS-insoluble, formic-acid-extractable proteins were found in the residual cell wall material. Two major proteins (12·8 and 14·0 kDa) were extracted with formic acid from fractured aerial and submerged conidia but not from blastospores. Oxidation of the sample extracted by formic acid resulted in a single protein band (15·4 kDa) as judged by SDS-PAGE. Antibodies against this cell wall protein (cwp1) did not cross-react with cell wall extracts from the entomopathogenic deuteromycetous fungi Verticillium lecanii or Western blot analysis of two-dimensional gels revealed at least three acidic isoforms (pl 4·0-4·8) of cwp1. Immunohistological studies revealed that the cwp1 was primarily localized in the cell wall of aerial and submerged conidia but not in blastospores, Immunolocalization was possible only if the conidia were previously boiled in 5% (v/v) β-mercaptoethanol. The N-terminal sequence of cwp1 showed no similarities with other published sequences. Our results suggest that at least two major species of SDS-insoluble, formic-acid-extractable proteins exist in cell walls of or submerged conidia; one is the hydrophobin which occurs in the outermost rodlet layer and the other, cwp1, occurs primarily next to the rodlet layer.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-5-1075
1995-05-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/5/mic-141-5-1075.html?itemId=/content/journal/micro/10.1099/13500872-141-5-1075&mimeType=html&fmt=ahah

References

  1. Bell-Peterson D., Dunlap J.C., Loros J.J. 1992; The Neurospora circadian clock-controlled gene, ccg-2, is allelic to eas and encodes a fungal hydrophobin required for formation of the conidial rodlet layer.. Genes & Dev 6:2382–2394
    [Google Scholar]
  2. Bidochka M.J., Pfeifer T.A., Khachatourians G.G. 1987; Development of the entomopathogenic fungus Beauveria bassiana in liquid cultures.. Mycopathologia 99:77–83
    [Google Scholar]
  3. Bidochka M.J., St Leger R.J., Joshi L., Roberts D.W. 1995; The rodlet layer from aerial and submerged conidia of the entomopathogenic fungus Beauveria bassiana contains hydrophobin.. Mycol Res (in press)
    [Google Scholar]
  4. Boucias D.G., Pendland J.C., Latge J.P. 1988; Nonspecific factors involved in attachment of entomopathogenic Deutero- mycetes to host insect cuticle.. Appl Environ Microbiol 54:1795–1805
    [Google Scholar]
  5. Capaldi R.A., Vanderkooi G. 1972; The low polarity of many membrane proteins.. Proc Natl Acad Sci USA 69:930–932
    [Google Scholar]
  6. Cole G.T., Takashi S., Kasai R., Yokoyama T., Nozawa Y. 1979; Surface ultrastructure and chemical composition of the cell walls of conidial fungi.. Exp Mycol 3:132–156
    [Google Scholar]
  7. Hegedus D.D., Bidochka M.J., Khachatourians G.G. 1990; Beauveria bassiana submerged conidia production in a defined medium containing chitin, two hexosamines or glucose.. Appl Microbiol Biotechnol 33:641–647
    [Google Scholar]
  8. Hegedus D.D., Bidochka M.J., Miranpuri G.S., Khachatourians G.G. 1992; A comparison of the virulence, stability and cell-wall surface characteristics of three spore types produced by the entomopathogenic fungus Beauveria bassiana.. Appl Microbiol Biotechnol 36:785–789
    [Google Scholar]
  9. Henrikson R.L., Meredith S.C. 1984; Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate.. Anal Biochem 136:65–74
    [Google Scholar]
  10. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  11. Lauter F.-R., Russo V.E.A., Yanofsky C. 1992; Development and light regulation of eas, the structural gene for the rodlet protein of Neurospora.. Genes & Dev 6:2373–2381
    [Google Scholar]
  12. Moos M. Jr Nguyen N.Y., Liu T.-Y. 1988; Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support.. J Biol Chem 263:6005–6008
    [Google Scholar]
  13. Reddy S.R.R., Wyatt G.R. 1967; Incorporation of uridine and leucine in vitro by cecropia silkmoth wing epidermis during diapause and development.. J Insect Physiol 13:981–994
    [Google Scholar]
  14. Stringer M.A., Timberlake W.E. 1993; Cerato-ulmin, a toxin involved in dutch elm disease, is a fungal hydrophobin.. Plant Cell 5:145–146
    [Google Scholar]
  15. Stringer M.A., Dean R.A., Sewall T.C., Timberlake W.E. 1992; Rodletless, a new Aspergillus developmental mutant induced by directed gene inactivation.. Genes & Dev 5:1161–1171
    [Google Scholar]
  16. Thomas K.C., Khachatourians G.G., Ingledew W.M. 1987; Production and properties of Beauveria bassiana conidia in submerged culture.. Can J Microbiol 33:12–20
    [Google Scholar]
  17. Tiedt L.R. 1993; An electron microscope study of conidiogenesis and wall formation of conidia of Aspergillus niger.. Mycol Res 97:1459–1462
    [Google Scholar]
  18. de Vries O.M.H., Fekkes M.P., Wosten H.A.B., Wessels J.G.H. 1993; Insoluble hydrophobin complexes in the walls of Schiypphyllum commune and other filamentous fungi.. Arch Microbiol 159:330–335
    [Google Scholar]
  19. Wessels J.G.H. 1992; Gene expression during fruiting in Schispphyllum commune.. Mycol Res 96:609–620
    [Google Scholar]
  20. Wessels J.G.H., de Vries O.M.H., Ásgeirsdóttir S.A., Springer J. 1991; The thn mutation of Schiophyllum commune, which suppresses formation of aerial hypae, affects expression of the Sc3 hydrophobin gene.. J Gen Microbiol 137:2439–2445
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-5-1075
Loading
/content/journal/micro/10.1099/13500872-141-5-1075
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