Bni5-mediated recruitment of Myo1 to the division site before cytokinesis endows the AMR robustness against perturbation during cytokinesis. (A) The Bni5–Myo1 interaction is essential for high levels of Myo1 accumulation at the bud neck prior to cytokinesis. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Asterisk indicates variants of Myo1. Strains used: YEF11441 (MYO1-GFP mScarlet-TUB1), YEF11442 (myo1-mTD1Δ-GFP mScarlet-TUB1), and YEF11443 (bni5Δ MYO1-GFP mScarlet-TUB1). (B) Disruption of F-actin compromises AMR constriction in bni5Δ cells. AMR (Myo1-GFP) constriction in the presence of moderate (50 µM) or high (200 µM) concentrations of LatA was scored based on the indicated categories from time-lapse series taken with 2-min intervals. n denotes the number of cells analyzed per strain. Strains used: YEF11385 (MYO1-GFP mScarlet-TUB1) and YEF11443. (C and D) Synthetic effects of bni5Δ and Myo1 tail truncation on Myo1 accumulation at the bud neck at 23°C and 37°C. Left: Accumulation kinetics of FL and tail-truncated Myo1 at the bud neck in WT and bni5Δ background were quantified from time-lapse series taken at 23°C (C) or 37°C (D) with 1.5-min intervals. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Right: Montages of indicated proteins at the bud neck were created from selected frames. Strains used: YEF11529 (GFP-MYO1 mScarlet-TUB1), YEF11524 [GFP-myo1(1-1797) mScarlet-TUB1], YEF11601 (bni5Δ GFP-MYO1 mScarlet-TUB1), and YEF11530 [bni5Δ GFP-myo1(1-1797) mScarlet-TUB1)]. (E) Quantification of AMR constriction phenotype in the indicated strains, based on imaging data from C and D. n denotes the number of cells analyzed per strain. Asterisk indicates variants of Myo1. (F) Cell morphology defects in bni5Δ and myo1 tail-truncation mutants. Cells were cultured in YM-1 medium at 25°C or 37°C for 24 h before imaging. Strains used are the same as in C and D. (G) Representative images of GFP-Myo1 and GFP-Myo1-1797 in homozygous and hemizygous diploid strains. Strains used: YEF6611 (GFP-MYO1/GFP-MYO1), YEF12297 (GFP-MYO1/myo1Δ), YEF6608 [GFP-myo1(1-1797)/GFP-myo1(1-1797)], and YEF12293 [GFP-myo1(1-1797)/myo1Δ]. (H) Quantification of GFP-Myo1 and GFP-Myo1-1797 fluorescence intensities from images in G. Green squares, brown bars, and gray bars represent individual data points, mean values, and SD, respectively. Black asterisk indicates variants of Myo1, and red asterisks indicate P < 0.01. P values were determined using a two-sided Mann–Whitney U test. (I and J) Cell morphology of the GFP-Myo1 and GFP-Myo1-1797 homozygous and hemizygous strains. (I) Representative images of cells of indicated strains were acquired as F. (J) Quantification of cell morphology. n denotes the number of cells analyzed per strain. Strains used are the same as in G.
Figure 8.

Bni5-mediated recruitment of Myo1 to the division site before cytokinesis endows the AMR robustness against perturbation during cytokinesis. (A) The Bni5–Myo1 interaction is essential for high levels of Myo1 accumulation at the bud neck prior to cytokinesis. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Asterisk indicates variants of Myo1. Strains used: YEF11441 (MYO1-GFP mScarlet-TUB1), YEF11442 (myo1-mTD1Δ-GFP mScarlet-TUB1), and YEF11443 (bni5Δ MYO1-GFP mScarlet-TUB1). (B) Disruption of F-actin compromises AMR constriction in bni5Δ cells. AMR (Myo1-GFP) constriction in the presence of moderate (50 µM) or high (200 µM) concentrations of LatA was scored based on the indicated categories from time-lapse series taken with 2-min intervals. n denotes the number of cells analyzed per strain. Strains used: YEF11385 (MYO1-GFP mScarlet-TUB1) and YEF11443. (C and D) Synthetic effects of bni5Δ and Myo1 tail truncation on Myo1 accumulation at the bud neck at 23°C and 37°C. Left: Accumulation kinetics of FL and tail-truncated Myo1 at the bud neck in WT and bni5Δ background were quantified from time-lapse series taken at 23°C (C) or 37°C (D) with 1.5-min intervals. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Right: Montages of indicated proteins at the bud neck were created from selected frames. Strains used: YEF11529 (GFP-MYO1 mScarlet-TUB1), YEF11524 [GFP-myo1(1-1797) mScarlet-TUB1], YEF11601 (bni5Δ GFP-MYO1 mScarlet-TUB1), and YEF11530 [bni5Δ GFP-myo1(1-1797) mScarlet-TUB1)]. (E) Quantification of AMR constriction phenotype in the indicated strains, based on imaging data from C and D. n denotes the number of cells analyzed per strain. Asterisk indicates variants of Myo1. (F) Cell morphology defects in bni5Δ and myo1 tail-truncation mutants. Cells were cultured in YM-1 medium at 25°C or 37°C for 24 h before imaging. Strains used are the same as in C and D. (G) Representative images of GFP-Myo1 and GFP-Myo1-1797 in homozygous and hemizygous diploid strains. Strains used: YEF6611 (GFP-MYO1/GFP-MYO1), YEF12297 (GFP-MYO1/myo1Δ), YEF6608 [GFP-myo1(1-1797)/GFP-myo1(1-1797)], and YEF12293 [GFP-myo1(1-1797)/myo1Δ]. (H) Quantification of GFP-Myo1 and GFP-Myo1-1797 fluorescence intensities from images in G. Green squares, brown bars, and gray bars represent individual data points, mean values, and SD, respectively. Black asterisk indicates variants of Myo1, and red asterisks indicate P < 0.01. P values were determined using a two-sided Mann–Whitney U test. (I and J) Cell morphology of the GFP-Myo1 and GFP-Myo1-1797 homozygous and hemizygous strains. (I) Representative images of cells of indicated strains were acquired as F. (J) Quantification of cell morphology. n denotes the number of cells analyzed per strain. Strains used are the same as in G.

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