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Vol. 14, Issue 10, 4140-4154, October 2003
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Applied Microbiology, Biozentrum, Universitaet Basel, 4056 Basel, Switzerland
Submitted March 22, 2003;
Revised June 17, 2003;
Accepted June 17, 2003
Monitoring Editor: David Drubin
We used actin staining and videomicroscopy to analyze the development from a spore to a young mycelium in the filamentous ascomycete Ashbya gossypii. The development starts with an initial isotropic growth phase followed by the emergence of germ tubes. The initial tip growth speed of 6–10 µm/h increases during early stages of development. This increase is transiently interrupted in response to the establishment of lateral branches or septa. The hyphal tip growth speed finally reaches a maximum of up to 200 µm/h, and the tips of these mature hyphae have the ability to split into two equally fast-growing hyphae. A search for A. gossypii homologs of polarisome components of the yeast Saccharomyces cerevisiae revealed a remarkable size difference between Spa2p of both organisms, with AgSpa2p being double as long as ScSpa2p due to an extended internal domain. AgSpa2 colocalizes with sites of polarized actin. Using time-lapse videomicroscopy, we show that AgSpa2p-GFP polarization is established at sites of branch initiation and then permanently maintained at hyphal tips. Polarization at sites of septation is transient. During apical branching the existing AgSpa2p-GFP polarization is symmetrically divided. To investigate the function of AgSpa2p, we generated two AgSPA2 mutants, a partial deletion of the internal domain alone, and a complete deletion. The mutations had an impact on the maximal hyphal tip growth speed, on the hyphal diameter, and on the branching pattern. We suggest that AgSpa2p is required for the determination of the area of growth at the hyphal tip and that the extended internal domain plays an important role in this process.
Online version of this article contains video material for some figures. Online version is available at www.molbiolcell.org.
Present address: Unité Biologie et Pathogénicité fongiques, Institut Pasteur, 25-28, Rue du Docteur Roux, 75724 Paris Cedex 15, France
Present address: Duke Center for Genome Technology, Duke University Medical Center, Box 3568, Durham, NC 27710-3568.
Corresponding author. E-mail address: peter.philippsen{at}unibas.ch.
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