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Eukaryotic Cell, December 2004, p. 1574-1588, Vol. 3, No. 6
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.6.1574-1588.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Deletion of the Dynein Heavy-Chain Gene DYN1 Leads to Aberrant Nuclear Positioning and Defective Hyphal Development in Candida albicans

R. Martin, A. Walther, and J. Wendland*

Junior Research Group, Growth-Control of Fungal Pathogens, Hans-Knöll Institute for Natural Products Research and Department of Microbiology, Friedrich-Schiller University, Hans-Knoell, Germany

Received 10 March 2004/ Accepted 30 August 2004

Cytoplasmic dynein is a microtubule-associated minus-end-directed motor protein. CaDYN1 encodes the single dynein heavy-chain gene of Candida albicans. The open reading frames of both alleles of CaDYN1 were completely deleted via a PCR-based approach. Cadyn1 mutants are viable but grow more slowly than the wild type. In vivo time-lapse microscopy was used to compare growth of wild-type (SC5314) and dyn1 mutant strains during yeast growth and after hyphal induction. During yeast-like growth, Cadyn1 strains formed chains of cells. Chromosomal TUB1-GFP and HHF1-GFP alleles were used both in wild-type and mutant strains to monitor the orientation of mitotic spindles and nuclear positioning in C. albicans. In vivo fluorescence time-lapse analyses with HHF1-GFP over several generations indicated defects in dyn1 cells in the realignment of spindles with the mother-daughter axis of yeast cells compared to that of the wild type. Mitosis in the dyn1 mutant, in contrast to that of wild-type yeast cells, was very frequently completed in the mother cells. Nevertheless, daughter nuclei were faithfully transported into the daughter cells, resulting in only a small number of multinucleate cells. Cadyn1 mutant strains responded to hypha-inducing media containing L-proline or serum with initial germ tube formation. Elongation of the hyphal tubes eventually came to a halt, and these tubes showed a defect in the tipward localization of nuclei. Using a heterozygous DYN1/dyn1 strain in which the remaining copy was controlled by the regulatable MAL2 promoter, we could switch between wild-type and mutant phenotypes depending on the carbon source, indicating that the observed mutant phenotypes were solely due to deletion of DYN1.

* Corresponding author. Mailing address: Hans-Knoell Institute for Natural Products Research e.V. and Friedrich-Schiller-University Department of Microbiology, Hans-Knoell Str.2, D-07745 Jena, Germany. Phone: 49-3641-65-7639. Fax: 49-3641-65-6620. E-mail: juergen.wendland{at}uni-jena.de.

Eukaryotic Cell, December 2004, p. 1574-1588, Vol. 3, No. 6
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.6.1574-1588.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sherwood, R. K., Bennett, R. J. (2008). Microtubule Motor Protein Kar3 Is Required for Normal Mitotic Division and Morphogenesis in Candida albicans. Eukaryot Cell 7: 1460-1474 [Abstract] [Full Text]  
  • Finley, K. R., Bouchonville, K. J., Quick, A., Berman, J. (2008). Dynein-dependent nuclear dynamics affect morphogenesis in Candida albicans by means of the Bub2p spindle checkpoint. J. Cell Sci. 121: 466-476 [Abstract] [Full Text]  
  • Dunkler, A., Wendland, J. (2007). Candida albicans Rho-Type GTPase-Encoding Genes Required for Polarized Cell Growth and Cell Separation. Eukaryot Cell 6: 844-854 [Abstract] [Full Text]  
  • Finley, K. R., Berman, J. (2005). Microtubules in Candida albicans Hyphae Drive Nuclear Dynamics and Connect Cell Cycle Progression to Morphogenesis. Eukaryot Cell 4: 1697-1711 [Abstract] [Full Text]  


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