Journal: Genetics

Article Title: Aspergillus SUMOylation mutants exhibit chromosome segregation defects including chromatin bridges

doi: 10.1093/genetics/iyad169

Figure Lengend Snippet: Chromatin bridges are surrounded by nuclear envelope with nuclear pore complexes. a) Images of Nup49-GFP (Nup49) and Histone-H1-mCherry (Histone-H1) in a wild-type control and the ΔsumO mutant. We have observed at least 20 bridges in the ΔsumO mutant, as evidenced from the Histone-H1 signals, and all contain Nup49-GFP (Nup49) signals on the bridges. b) Images of Nup96-GFP (Nup96), NLS-Ds-Red (NLS), and Histone-H1-mCherry (Histone-H1) in a wild-type control and the ΔsumO mutant. Note that the NLS and Histone-H1 signals on the right panel of the ΔsumO images have been enhanced to show the bridge. All bridges showed Nup96 signals (n > 20). Bar: 10 µm.

Article Snippet: An ET-EGFP/mCherry Dual Multiband Filter Set w/BX3 cube (purchased from Olympus Inc.) was used.

Techniques: Mutagenesis

Journal: Genetics

Article Title: Aspergillus SUMOylation mutants exhibit chromosome segregation defects including chromatin bridges

doi: 10.1093/genetics/iyad169

Figure Lengend Snippet: Loss of SUMOylation does not cause early endosomes to accumulate abnormally at the hyphal tip. a) Distribution of mCherry-Rab5A-labeled early endosomes in the wild type, the ubaBQ247* mutant and the ΔsumO mutant. The hyphal tip is indicated by a arrowhead. As a control exhibiting an abnormal hyphal-tip accumulation of mCherry-Rab5A signals, we used a recently studied kinesin-1 mutant, kinAK895* (Qiu, Zhang, and Xiang 2023). The ubaBQ247* and ΔsumO mutants show a normal distribution of early endosomes, as compared to the wild type and the kinAK895* mutant. Bar: 10 µm. b) Line scans of mCherry-Rab5A (early endosomes) fluorescence intensity in the wild type, the ΔsumO mutant and the kinAK895* mutant. All values are relative to the mean value for wild type at position 0, which is set as 1. XY graphs with mean (solid lines) and standard error of the mean (SEM, shading) were generated by Prism 9. The intensity of mCherry-Rab5A near the hyphal tip (between 0.455 and 3.185 µm from hyphal tip with 0.065 µm as intervals) was significantly higher in the kinAK895* mutant than that in both the wild-type and the ΔsumO mutant (P < 0.0001, 2-way ANOVA with Tukey's multiple comparisons test, n = 31 hyphae for wild type, n = 31 hyphae for the ΔsumO mutant, and n = 29 hyphae for the kinAK895* mutant). The overall intensity of mCherry-Rab5A appeared higher in the ΔsumO mutant than in the wild type along the hyphae, but the differences were not statistically significant (P > 0.05 for all the points from 0 to 10.595 µm from the hyphal tip).

Article Snippet: An ET-EGFP/mCherry Dual Multiband Filter Set w/BX3 cube (purchased from Olympus Inc.) was used.

Techniques: Labeling, Mutagenesis, Fluorescence, Generated

Journal: Genetics

Article Title: Aspergillus SUMOylation mutants exhibit chromosome segregation defects including chromatin bridges

doi: 10.1093/genetics/iyad169

Figure Lengend Snippet: SUMOylation deficiency causes a severe defect in the first nuclear division at a higher temperature of 42°C. a) Nuclei labeled with Histone-H1-GFP in wild type, the ΔubaB single mutant, the nimT23 single mutant, and the nimT23, ΔubaB double mutant. Images were taken after a 6-h incubation at the restrictive temperature of 42°C (42°C) or after shifting the cells to 25°C for 0.5–1 h (42–25°C). Bar: 5 µm. b) A quantitative analysis on the percent of germ tubes (42–25°C) containing a chromatin bridge (ordinary 1-way ANOVA with Tukey's multiple comparisons test). Scatter plots with mean and SD values were generated by Prism 9. c) A quantitative analysis on the percent of germ tubes (42–25°C) containing an abnormally shaped nucleus (ordinary 1-way ANOVA with Tukey's multiple comparisons test). Scatter plots with mean and SD values were generated by Prism 9. For b andc, data were from 3 experiments. For each experiment, at least 40 germ tubes were counted for the ΔubaB single mutant, the nimT23 single mutant, and the nimT23, ΔubaB double mutant, and at least 24 germ tubes were counted for the wild-type strain. The total germ tubes counted were 109 for wild type, 229 for ΔubaB, 142 for nimT23, and 234 for ΔubaB, nimT23. d) Nuclei labeled with Nup49-GFP (top) and Histone-H1-mCherry (bottom) in wild type and the ΔsumO mutant grown at 42°C for 7 h.

Article Snippet: An ET-EGFP/mCherry Dual Multiband Filter Set w/BX3 cube (purchased from Olympus Inc.) was used.

Techniques: Labeling, Mutagenesis, Incubation, Generated