Identification and Characterization of Aspergillus nidulans Mutants Defective in Cytokinesis

INVESTIGATIONS

Identification and Characterization of Aspergillus nidulans Mutants Defective in Cytokinesis

S. D. Harris, J. L. Morrell and J. E. Hamer
Department of Biological Sciences, Hansen Life Science Research Building, Purdue University, West Lafayette, Indiana 47907-1392

Filamentous fungi undergo cytokinesis by forming crosswalls termed septa. Here, we describe the genetic and physiological controls governing septation in Aspergillus nidulans. Germinating conidia do not form septa until the completion of their third nuclear division. The first septum is invariantly positioned at the basal end of the germ tube. Block-and-release experiments of nuclear division with benomyl or hydroxyurea, and analysis of various nuclear division mutants demonstrated that septum formation is dependent upon the third mitotic division. Block-and-release experiments with cytochalasin A and the localization of actin in germlings by indirect immunofluorescence showed that actin participated in septum formation. In addition to being concentrated at the growing hyphal tips, a band of actin was also apparent at the site of septum formation. Previous genetic analysis in A. nidulans identified four genes involved in septation (sepA-D). We have screened a new collection of temperature sensitive (ts) mutants of A. nidulans for strains that failed to form septa at the restrictive temperature but were able to complete early nuclear divisions. We identified five new genes designated sepE, G, H, I and J, along with one additional allele of a previously identified septation gene. On the basis of temperature shift experiments, nuclear counts and cell morphology, we sorted these cytokinesis mutants into three phenotypic classes. Interestingly, one class of mutants fails to form septa and fails to progress past the third nuclear division. This class of mutants suggests the existence of a regulatory mechanism in A. nidulans that ensures the continuation of nuclear division following the initiation of cytokinesis.

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				A. Virag and S. D. Harris Functional Characterization of Aspergillus nidulans Homologues of Saccharomyces cerevisiae Spa2 and Bud6 Eukaryot. Cell, June 1, 2006; 5(6): 881 - 895. [Abstract] [Full Text] [PDF]

				J.-M. Kim, L. Lu, R. Shao, J. Chin, and B. Liu Isolation of Mutations That Bypass the Requirement of the Septation Initiation Network for Septum Formation and Conidiation in Aspergillus nidulans Genetics, June 1, 2006; 173(2): 685 - 696. [Abstract] [Full Text] [PDF]

				T. W. Hill, D. M. Loprete, M. Momany, Y. Ha, L. M. Harsch, J. A. Livesay, A. Mirchandani, J. J. Murdock, M. J. Vaughan, and M. B. Watt Isolation of cell wall mutants in Aspergillus nidulans by screening for hypersensitivity to Calcofluor White. Mycologia, May 1, 2006; 98(3): 399 - 409. [Abstract] [Full Text] [PDF]

				I. Malavazi, C. P. Semighini, M. R. von Zeska Kress, S. D. Harris, and G. H. Goldman Regulation of Hyphal Morphogenesis and the DNA Damage Response by the Aspergillus nidulans ATM Homolog AtmA Genetics, May 1, 2006; 173(1): 99 - 109. [Abstract] [Full Text] [PDF]

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				S. Li, L. Du, G. Yuen, and S. D. Harris Distinct Ceramide Synthases Regulate Polarized Growth in the Filamentous Fungus Aspergillus nidulans Mol. Biol. Cell, March 1, 2006; 17(3): 1218 - 1227. [Abstract] [Full Text] [PDF]

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				A. Beauvais, D. Maubon, S. Park, W. Morelle, M. Tanguy, M. Huerre, D. S. Perlin, and J. P. Latge Two {alpha}(1-3) Glucan Synthases with Different Functions in Aspergillus fumigatus Appl. Envir. Microbiol., March 1, 2005; 71(3): 1531 - 1538. [Abstract] [Full Text] [PDF]

				S. E. Gygax, C. P. Semighini, G. H. Goldman, and S. D. Harris SepBCTF4 Is Required for the Formation of DNA-Damage-Induced UvsCRAD51 Foci in Aspergillus nidulans Genetics, March 1, 2005; 169(3): 1391 - 1402. [Abstract] [Full Text] [PDF]

				J. R. Davies, A. H. Osmani, C. P. C. De Souza, C. Bachewich, and S. A. Osmani Potential Link between the NIMA Mitotic Kinase and Nuclear Membrane Fission during Mitotic Exit in Aspergillus nidulans Eukaryot. Cell, December 1, 2004; 3(6): 1433 - 1444. [Abstract] [Full Text] [PDF]

				K. S. Bruno, F. Tenjo, L. Li, J. E. Hamer, and J.-R. Xu Cellular Localization and Role of Kinase Activity of PMK1 in Magnaporthe grisea Eukaryot. Cell, December 1, 2004; 3(6): 1525 - 1532. [Abstract] [Full Text] [PDF]

				M. Martin-Udiroz, M. P. Madrid, and M. I. G. Roncero Role of chitin synthase genes in Fusarium oxysporum Microbiology, October 1, 2004; 150(10): 3175 - 3187. [Abstract] [Full Text] [PDF]

				M. Momany, R. Lindsey, T. W. Hill, E. A. Richardson, C. Momany, M. Pedreira, G. M. Guest, J. F. Fisher, R. B. Hessler, and K. A. Roberts The Aspergillus fumigatus cell wall is organized in domains that are remodelled during polarity establishment Microbiology, October 1, 2004; 150(10): 3261 - 3268. [Abstract] [Full Text] [PDF]

				C. L. Pearson, K. Xu, K. E. Sharpless, and S. D. Harris MesA, a Novel Fungal Protein Required for the Stabilization of Polarity Axes in Aspergillus nidulans Mol. Biol. Cell, August 1, 2004; 15(8): 3658 - 3672. [Abstract] [Full Text] [PDF]

				J. D. Joseph, S. N. Daigle, and A. R. Means PINA Is Essential for Growth and Positively Influences NIMA Function in Aspergillus nidulans J. Biol. Chem., July 30, 2004; 279(31): 32373 - 32384. [Abstract] [Full Text] [PDF]

				T. Oka, T. Hamaguchi, Y. Sameshima, M. Goto, and K. Furukawa Molecular characterization of protein O-mannosyltransferase and its involvement in cell-wall synthesis in Aspergillus nidulans Microbiology, June 1, 2004; 150(6): 1973 - 1982. [Abstract] [Full Text] [PDF]