Summary
A monoclonal antibody was used to localize γ-tubulin in hyphal tip cells of the chytridiomycete fungus Allomyces macrogynus, and its distribution determined with standard epifluorescence and laser scanning confocal microscopy. The results demonstrate that γ-tubulin is a component of the Spitzenkörper and centrosomes. Immunoblot analysis of total soluble protein extracts separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis identified a single 56 kDa γ-tubulin-related polypeptide. Localization of γ-tubulin to the Spitzenkörper of A. macrogynus provides evidence that the Spitzenkörper in this fungus functions as a microtubule-organizing center.
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Abbreviations
- BSA:
-
bovine serum albumin
- DAPI:
-
4′,6-diamidino-2-phenylindole
- DIC:
-
differential interference contrast
- LSCM:
-
laser scanning confocal microscopy
- MTOCs:
-
microtubule-organizing centers
- PBS:
-
phosphate-buffered saline
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- SPB:
-
spindle pole body
- YpSs:
-
yeast extract-inorganic phosphate-soluble starch
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McDaniel, D.P., Roberson, R.W. γ-Tubulin is a component of the Spitzenkörper and centrosomes in hyphal-tip cells of Allomyces macrogynus . Protoplasma 203, 118–123 (1998). https://doi.org/10.1007/BF01280594
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DOI: https://doi.org/10.1007/BF01280594