Figure 1.
Colony morphology of Aspergillus nidulans wild type (WT) strain complemented with wild type AfUgmA (WC), single residue AfUgmA mutants (F66A, H63N, R182K, R182A, R327K, R327A) and AnugmAΔ strains, grown on complete medium at 30°C for 3 d.
The colour difference between WT and WC strains was due to slightly different ages of culture. See Figure S1 in File S1 for a direct comparison of the spore colours of these two strains.
Table 1.
Colony and hyphal characteristicsa of wild type, and AnugmA▵ strains complemented with wild type AfUgmA, single residue AfUgmA mutants and deletion strains grown on CM.
Table 2.
Aspergillus nidulans wild type and chimaera cell wall carbohydratesa, antifungal drug sensitivityac, and relative UGM activityb.
Figure 2.
Surface adhesion of Aspergillus nidulans strains to fluorescent latex beads.
Wild type (WT), strain complemented with wild type AfUgmA (WC), single residue AfUgmA mutants (F66A, H63N and R327A) and AnugmAΔ strains. Bar = 20 µm is for all images.
Figure 3.
In vivo distribution of GFP-tagged AfUgmA in Aspergillus nidulans.
A. Wild type complemented (WC) and single residue mutants (H63N, R182A and R327A) have comparable AfUgmA-GFP distribution. The single residue mutants have the ugmAΔ hyphal morphology. Bar = 20 µm for all images. B. Confirmation of AfUgmA-GFP fusion protein distribution by Western blot. Total protein was extracted from A. nidulans wild type (WT; AAE1) and GFP-tagged AfUgmA strains (R182A-GFP, R327A-GFP, H63N-GFP, AnUgmA::AfUgm-GFP). Total Proteins (15 µg/lane) were separated on 10% SDS-PAGE and immunoblotted with an anti-GFP antibody.
Figure 4.
Localization of Galactofuranose (Galf) in Aspergillus nidulans Cell wall.
A. Galf immunolocalization. B. Immunofluorescence quantification of Galf using confocal system software (see Methods). Error bar shows standard error. Aspergillus nidulans wild type (WT), wild type AfUgmA-complemented (WC), mutated AfUgmA (as listed), and AnugmAΔ. Bar = 10 μm for all images.
Figure 5.
Localization of Alpha-glucan in Aspergillus nidulans Cell wall.
A. Alpha-glucan immunolocalization. B. Immunofluorescence quantification of Alpha-glucan using confocal system software (see Methods). Error bar shows standard error. Aspergillus nidulans wild type (WT), wild type AfUgmA-complemented (WC), mutated AfUgmA (as listed), and AnugmAΔ. Bar = 10 μm for all images.
Figure 6.
Localization of Beta-glucan in Aspergillus nidulans Cell wall.
A. Beta-glucan immunolocalization. B. Immunofluorescence quantification of Beta-glucan using confocal system software (see Methods). Error bar shows standard error. Aspergillus nidulans wild type (WT), wild type AfUgmA-complemented (WC), mutated AfUgmA (as listed), and AnugmAΔ. Bar = 10 μm for all images.