Observation of EGFP-visualized nuclei and distribution of vacuoles in Aspergillus oryzae arpA null mutant

• Published in: FEMS microbiology letters Vol. 206; no. 1; pp. 57 - 61
• Main Authors: Maruyama, J, Nakajima, H, Kitamoto, K
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.01.2002; Blackwell; Oxford University Press
• Subjects: Actin; Actins - genetics; Actins - metabolism; Actin‐related protein; Arp1 protein; arpA gene; Aspergillus nidulans; Aspergillus oryzae; Aspergillus oryzae - genetics; Aspergillus oryzae - metabolism; Aspergillus oryzae - ultrastructure; Biological and medical sciences; Cell migration; Cell Nucleus - metabolism; Cell Nucleus - ultrastructure; Conidia; conidiophores; Dynactin Complex; EGFP; Fundamental and applied biological sciences. Psychology; Fungal Proteins; Fusion protein; Gene Deletion; genes; Genes, Fungal; Green Fluorescent Proteins; Growth rate; Histone H2B; histones; Histones - genetics; Histones - metabolism; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Microbiology; Microtubule-Associated Proteins - metabolism; Morphology; Morphology, structure, chemical composition; Mutants; Mycology; Nuclear transport; Organelles; phenotype; Phenotypes; Proteins; Recombinant Fusion Proteins; Transformation, Genetic; Vacuole; Vacuoles; Vacuoles - metabolism; Vacuoles - ultrastructure; Dynactin complex; Actin-related protein; EGFP; Vacuole; Histone H2B; Growth rate; Fungi; Aspergillus oryzae; Gene; Actin; Aspergillus nidulans; Morphology; Green fluorescent protein; Fungi Imperfecti; Mutation; Fusion protein; Thallophyta
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.2002.tb10986.x

The arpA gene encoding Arp1 (actin-related protein) was previously cloned and characterized from Aspergillus oryzae. Phenotypes of the arpA null mutant indicate its requirement for normal nuclear distribution and morphology of conidiophores. In this study, we further characterized the function of the arpA gene in distribution of organelles. For further analysis of nuclear migration in living cells, an expression system consisting of a fusion protein of Aspergillus nidulans histone H2B and EGFP (H2B::EGFP) was used. This demonstrated diminished hyphal-tip growth rate and inefficient nuclear transport to apical regions in the arpA null mutant. Expression of H2B::EGFP also revealed an increase in the nuclear number of each conidium in the arpA null mutant, implicating a role for the arpA gene in controlling the nuclear movement into conidia. Furthermore, staining of vacuoles of the arpA null mutant with CMAC (cell tracker blue) suggested that the arpA gene is required for proper vacuolar distribution in addition to its role in normal nuclear distribution.