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Microbiology 144 (1998), 45-53; DOI  10.1099/00221287-144-1-45
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Effect of cytochalasin A on apical growth, actin cytoskeleton organization and enzyme secretion in Aspergillus nidulans

Sara Torralba1, Marjatta Raudaskoski2, Ana María Pedregosa1 and Fernando Laborda1

Departament de Microbiología y Parasitología, Universidad de Alcalá de Henares, Carretera Madrid-Barcelona, Km 33, E-28871 Alcalá de Henares, Madrid, Spain
Division of Plant Physiology, Department of Biosciences, University of Helsinki, PO Box 56, Viikinkaari 9, FIN-00014, University of Helsinki, Finland

ABSTRACT

Summary: The role of actin in apical growth and enzyme secretion in the filamentous fungus Aspergillus nidulans was studied by treating the hyphae with cytochalasin A (CA), which inhibits actin polymerization. Indirect immunofluorescence microscopy revealed actin at the tips of main hyphae and branches, and at the sites of developing septa. CA inhibited the growth of the fungus and changed the growth pattern of hyphal tips from cylindrical tubes to spherical beads. The regions with swellings showed no actin fluorescence, and neither was actin seen in association with septa. After 4 h exposure, hyphae were able to resume the normal tip growth pattern in the presence of CA for a short period of time and new cylindrical hyphae, with actin fluorescence at the apex, emerged from the swollen tips. Later, the tips of the hyphae swelled again, which led to a beaded apperance. We also studied the effect of CA on the secretion of {alpha}- and β-galactosidase. {alpha}-Galactosidase is secreted into the culture medium, whereas β-galactosidase remains in the mycelium, with part of its activity bound to the cell wall. When A. nidulans mycelium was incubated in the presence of CA, a reduction in the secretion of {alpha}-galactosidase into the culture medium and a decrease in the {alpha}- and β-galactosidase activities bound to the cell wall was detected. However, the CA dose used for the hyphae did not modify the secretion of the enzymes from protoplasts. Results described here provide evidence that a polymerized actin cytoskeleton is required for normal apical growth, hyphal tip shape and polarized enzyme secretion in A. nidulans. Cytochalasin-induced disruptions of the actin cytoskeleton could result in the alterations of apical growth and inhibition of enzyme secretion observed by blocking secretory vesicle transport to the apex.

Author for correspondence: Sara Torralba. Fax: +34 1 8854621. e-mail: mpampp@microb.alcala.es

Keywords: actin, cytochalasin, Aspergillus nidulans, {alpha}-galactosidase, β-galactosidase




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