1. Cell Biology

Nim1-related kinases regulate septin organization and cytokinesis by modulating Hof1 at the cell division site

  1. Bindu Bhojappa
  2. Anubhav Dhar
  3. VT Bagyashree
  4. Jayanti Kumari
  5. Freya Cardozo
  6. Vaseef Rizvi
  7. Deepthi Guturu
  8. Saravanan Palani  Is a corresponding author
  1. Department of Biochemistry, Division of Biological Sciences, Indian Institute of Science, India
https://doi.org/10.7554/eLife.106366.3
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Cite this article
  1. Bindu Bhojappa
  2. Anubhav Dhar
  3. VT Bagyashree
  4. Jayanti Kumari
  5. Freya Cardozo
  6. Vaseef Rizvi
  7. Deepthi Guturu
  8. Saravanan Palani
(2026)
Nim1-related kinases regulate septin organization and cytokinesis by modulating Hof1 at the cell division site
eLife 14:RP106366.
https://doi.org/10.7554/eLife.106366.3
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14 figures, 3 tables and 4 additional files

Figures

Figure 1 with 5 supplements
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Septin organization and actomyosin ring (AMR) dynamics are defective in elm1Δ and gin4Δ cells.

(A) Domain-level architecture of Elm1, Gin4, Kcc4, and Hsl1 kinases. The KA1 domain represents the membrane-binding domain. (B) Representative time-lapse images showing Cdc3-mCherry dynamics in septin-kinase deletion strains, captured at 2-min time intervals (t0 = spindle breakpoint). (C) Normalized fluorescence intensity profile showing the temporal kinetics of Cdc3-mCherry in the indicated strains shown in (B) (wild-type: n = 41, elm1Δ: n = 29, gin4Δ: n = 33, hsl1Δ: n = 34, kcc4Δ: n = 36 cells). (D) Representative time-lapse images of Inn1-mNG constriction captured at 2-min time intervals in septin-kinase deletion strains (t0 = appearance of Inn1-mNG at the bud neck). (E) Quantification of the residence time of Inn1-mNG during the onset of cytokinesis in the indicated strains of (D).Kruskal–Wallis nonparametric test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × standard deviation (SD). (F) Normalized fluorescence intensity graph showing the kinetics of Inn1-mNG in the indicated strains of (D). A population of elm1Δ and gin4Δ cells exhibiting asymmetric constriction is included in the plot (wild-type: n = 21, elm1Δ: n = 13, gin4Δ: n = 23, hsl1Δ: n = 21, and kcc4Δ: n = 18 cells).

Figure 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
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Dynamics of septin-associated kinases exhibiting simultaneous recruitment during bud emergence and ordered disassembly during the HDR transition.

(A) Representative time-lapse images and normalized fluorescence intensity graph showing the temporal kinetics of Elm1-GFP during bud-emergence (t0) (n = 18 cells). (B) Representative time-lapse images and normalized fluorescence intensity graph showing the temporal kinetics of Gin4-GFP during bud-emergence (t0) (n = 16 cells). (C) Representative time-lapse images and normalized fluorescence intensity graph showing the temporal kinetics of Hsl1-GFP during bud-emergence (t0) (n = 18 cells). (D) Representative time-lapse images and normalized fluorescence intensity graph for temporal kinetics of Kcc4-GFP during bud-emergence (t0) (n = 16 cells). Images in panels A-D correspond to differential intensity contrast. (E) Representative bud neck montages of Elm1-GFP, Gin4-GFP, Hsl1-GFP, and Kcc4-GFP during HDR transition (t0 = Cdc3-mCherry splitting), DC* = differential contrast. (F) Quantitative analysis of raw fluorescence intensity showing the dynamics of septin kinases indicated in (E) during septin remodeling. The Cdc3-mCherry profile of the Kcc4 strain is plotted as a reference for septin remodeling dynamics during cytokinesis. (Elm1-GFP: n = 22, Gin4-GFP: n = 26, Hsl1-GFP: n = 17, and Kcc4-GFP: n = 22 cells). (G) Plot of normalized fluorescence intensity at the bud neck for the indicated strains shown in (E). The Cdc3-mCherry profile of the Kcc4 strain is plotted as a reference for septin remodeling.

Figure 1—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 1—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig1-figsupp1-data1-v1.xlsx
Figure 1—figure supplement 2
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Deletion of septin-associated kinases differentially perturbs cellular morphology and septin organization.

(A) Brightfield images showing the cellular morphological defects in elm1Δ, gin4Δ, hsl1Δ, and kcc4Δ cells at 23, 30, and 37°C temperatures. Scale bar: 5 µm. (B) Stacked column graphs showing the percentage of cells exhibiting normal and abnormal cell shape in the indicated strains shown in (A). One-way ANOVA with Tukey’s multiple-comparison test. Error bars indicate + SD from the mean (N = 3, n = 300 cells/strain). (C) Growth assay of the indicated strains shown in (A) at 23, 30, and 37°C. Representative images were captured after 48 hr of incubation at indicated temperatures. (D) Representative montage showing the temporal kinetics of Cdc3-mCherry at 1-min time intervals during mitotic spindle break (t0). Scale bar: 5 µm. (E) Images representing the normal and mislocalization of Cdc3-mCherry during bud emergence in the indicated strains shown in (B). White arrows depict mislocalized septins in elm1Δ and gin4Δ strains. Scale bar: 5 µm. (F) Normalized fluorescence intensity graph showing the kinetics of Cdc3-mCherry during cytokinesis in the represented strain shown in (D). (G) Stacked column graph showing the percentage of cells exhibiting normal localization and mislocalization of Cdc3-mCherry to the bud cortex in the indicated strains of (E) (wild-type: n = 45, elm1Δ: n = 48, gin4Δ: n = 41, hsl1Δ: n = 56, kcc4Δ: n = 50 cells).

Figure 1—figure supplement 2—source data 1

Numerical data underlying the quantitative analyses represented in Figure 1—figure supplement 2.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig1-figsupp2-data1-v1.xlsx
Figure 1—video 1
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Representative movies showing the recruitment of Elm1, Gin4, Hsl1, and Kcc4 during bud emergence and cytokinesis.

Scale bar: 5 µm.

Figure 1—video 2
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Representative movies showing the localization and dynamics of Cdc3-mCherry in wild-type, elm1Δ, gin4Δ, kcc4Δ, and hsl1Δ backgrounds.

Scale bar: 5 µm.

Figure 1—video 3
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Representative movies showing Inn1-mNG constriction in wild-type, elm1Δ, gin4Δ, kcc4Δ, and hsl1Δ backgrounds.

Scale bar: 5 µm.

Figure 2 with 3 supplements
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Gin4 regulates Hof1 organization and dynamics through interaction with the N-terminal membrane-binding F-BAR domain.

(A) Representative time-lapse images of Hof1-mNG in wild-type, elm1Δ, and gin4Δ cells imaged at 2-min time intervals (t0 = spindle breakpoint). (B) Quantification of the residence time of Hof1-mNG after spindle breakpoint in the indicated strains shown in (A).Kruskal–Wallis nonparametric test (**p < 0.01, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD. (C) Normalized fluorescence intensity profile for the temporal kinetics of Hof1-mNG during cytokinesis in the indicated strains shown in (A), (wild-type: n = 33, elm1Δ: n = 21, gin4Δ: n = 23 cells). (D) Representative images showing the organization of Hof1-mNG at the bud neck in the indicated strains shown in (A). Scale bar: 5 µm. (E) Bar graph showing the percentage of cells exhibiting misorganization of Hof1 at the cell division site in the indicated strains shown in (D) .One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean. p-values correspond to the population of cells exhibiting disorganized Hof1-mNG at the bud neck (N = 3, wild-type: n = 518, elm1Δ: n = 225, gin4Δ: n = 384 cells). (F) Representative images from the Yeast Two-Hybrid assay depicting the interaction of full-length Gin4, Gin4-KID, and Gin4-KA1 with full-length Hof1. (G) Representative Yeast Two-Hybrid images showing the interaction between Gin4-KA1 (1003–1142 aa) and the N-terminal F-BAR containing domain of Hof1 (1–350 aa). (H) Immunoblot showing the in vitro binding assay for the interaction between 6xHis-bdSUMO-Gin4KA1 and GST-Hof1F-BAR fragments. GST-tagged Hof1 fragment is immobilized on glutathione resin, and bound 6xHis-bdSUMO-tagged Gin4 fragments were analyzed by SDS–PAGE. Input (top) and pull-down fractions (middle) were probed with anti-His antibody, while GST-Hof1 fragments (bottom) were detected using anti-GST antibody, (+) present, (−) absent. (I) Proposed model depicting the possible interplay between Gin4 and Hof1 at the plasma membrane interface. Created with BioRender.com.

Figure 2—source data 1

Numerical data underlying the quantitative analyses represented in Figure 2.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig2-data1-v1.xlsx
Figure 2—source data 2

PDF containing the uncropped western blots corresponding to the in vitro binding assay shown in Figure 2H, with the relevant bands and regions clearly indicated.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig2-data2-v1.zip
Figure 2—source data 3

Original files of the full, uncropped, unedited western blots corresponding to the in vitro binding assay shown in Figure 2H.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig2-data3-v1.zip
Figure 2—figure supplement 1
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Gin4 regulates Hof1 organization and cell viability in a kinase-independent manner.

(A) Representative images showing the localization profile of Myo1-mNG in elm1Δ and gin4Δ cells. White arrows indicate the mislocalized Myo1 in the represented strains. Scale bar: 5 µm. (B) Stacked column graph showing the percentage of cells exhibiting normal localization and mislocalization of Myo1 in the indicated strains of (A) (wild-type: n = 26, elm1Δ: n = 29 and gin4Δ: n = 30 cells). (C) Representative time-lapse images of Chs2-mNG in wild-type, elm1Δ, and gin4Δ cells imaged at 2-min time intervals (t0 = appearance of Chs2 at the bud neck). (D) Quantification of the residence time of Chs2-mNG at the bud neck in indicated strains shown in (C) .Kruskal–Wallis nonparametric test (**p < 0.01, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD. (E) Plot of normalized fluorescence intensity of Chs2-mNG in indicated strains of (C). A population of elm1Δ and gin4Δ cells exhibiting asymmetric constriction is plotted in the graph (t0 = spindle breakpoint) (wild-type: n = 40, elm1Δ: n = 9, gin4Δ: n = 23 cells). (F) Representative images showing rescue of Hof1-misorganization at the cell division site upon expression of Gin4FL and Gin4KD constructs cloned under the endogenous promoter in gin4Δ cells. Scale bar: 5 µm. (G) Graph representing the percentage of cells exhibiting normal organization and misorganization of Hof1 rings in the indicated strains shown in (F).One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean. p-values correspond to the population of cells exhibiting disorganized Hof1 at the bud neck (N = 3, wild-type: n = 284, gin4Δ-Empty vector: n = 138, gin4Δ-Gin4FL: n = 148, gin4Δ-Gin4KD: n = 113 cells). (H) Representative images from the Yeast Two-Hybrid assay depicting the interaction between Gin4-KA1 domain and different domains of Hof1. (I) Spot assay showing rescue of synthetic lethality upon expression of Gin4FL and Gin4KD constructs cloned under the endogenous promoter in gin4Δ hof1Δ cells. Plates were incubated at 23°C and scanned after 72 hr.

Figure 2—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 2—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig2-figsupp1-data1-v1.xlsx
Figure 2—figure supplement 2
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Gin4-ka1Δ-GFP exhibits reduced localization to the bud neck prior to cytokinesis and recapitulates Hof1 defects observed in gin4Δ cells.

(A) Bud neck montages representing the temporal kinetics of Gin4-GFP in comparison with Gin4-ka1Δ-GFP (t0 = spindle breakpoint). (B) Plot depicting the raw fluorescence intensity profile of the indicated strains shown in (A) (Gin4-GFP: n = 29 and Gin4-ka1Δ-GFP: n = 29 cells). (C) Temporal kinetics graph showing normalized fluorescence intensity for the indicated strains shown in (A). (D) Representative montages showing the temporal kinetics of Hof1-mNG during cytokinesis at 1-min time intervals in gin4Δ and Gin4-ka1Δ cells (t0 = spindle breakpoint). (E) Quantification of the residence time of Hof1-mNG after spindle breakpoint in the indicated strains shown in (D).Kruskal–Wallis nonparametric test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD. (F) Temporal kinetics graph of the normalized fluorescence intensity of Hof1-mNG during cytokinesis in the indicated strains shown in (D) (wild-type: n = 30, gin4Δ: n = 29, and Gin4-ka1Δ: n = 30 cells).

Figure 2—figure supplement 2—source data 1

Numerical data underlying the quantitative analyses represented in Figure 2—figure supplement 2.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig2-figsupp2-data1-v1.xlsx
Figure 2—video 1
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Representative movies showing the localization and constriction of Hof1-mNG in wild-type, elm1Δ, and gin4Δ cells during the cell cycle.

Scale bar: 5 µm.

Figure 3
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Gin4 controls septin organization and AMR dynamics independently of its kinase activity.

(A) Representative images showing septin defects or mislocalization during bud emergence in gin4Δ cells and rescue of the mislocalization by Gin4FL-GFP and Gin4KD-GFP constructs expressed under pTEF promoter. White arrows indicate mislocalized Cdc3-mcherry in the represented strains. Scale bar: 5 µm. (B) Quantitation of septin mislocalization and its rescue in the indicated strains shown in (A) (wild-type: n = 38, gin4Δ: n = 32, gin4Δ-Gin4FL-GFP: n = 32, gin4Δ-Gin4KD-GFP: n = 36 cells). (C) Representative time-lapse images showing Inn1-mNG dynamics acquired at 2-min time intervals in gin4Δ cells expressing Gin4FL and Gin4KD constructs cloned under endogenous promoter (t0 = appearance of Inn1-mNG at the bud neck). (D) Plot of normalized fluorescence intensity of Inn1-mNG in indicated strains shown in (C). A population of gin4Δ-Empty vector cells exhibiting asymmetric constriction is plotted in the graph (t0 = spindle breakpoint) (wild-type: n = 38, gin4Δ-Empty vector: n = 21, gin4Δ-Gin4FL: n = 39, gin4Δ-Gin4KD: n = 31 cells). (E) Quantification of the residence time of Inn1-mNG during cytokinesis in indicated strains of (C).Kruskal–Wallis nonparametric test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD.

Figure 3—source data 1

Numerical data underlying the quantitative analyses represented in Figure 3.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig3-data1-v1.xlsx
Figure 4 with 1 supplement
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Artificial tethering of Gin4-GFP to the bud neck via Hsl1-GBP restores functionality in elm1Δ cells in a Gin4 kinase-independent manner.

(A) Schematic representation of GFP-GBP artificial tethering strategy, in which the protein of interest is targeted to the bud neck by C-terminal GFP tagging, while the binding partner is tagged with GBP. Created with BioRender.com. (B) Representative images showing artificial tethering of Gin4-GFP with Hsl1-GBP in elm1Δ cells. Scale bar: 5 µm. (C) Bar graph showing the percentage of cells exhibiting round morphology in the indicated strains shown in (B).One-way ANOVA with Tukey’s multiple-comparison test (**p < 0.01, ****p < 0.0001).Error bars indicate + SD from the mean (N = 3, wild-type: n = 407, elm1Δ: n = 508, elm1Δ-Hsl1-GBP: n = 400 cells). (D) Quantification of aspect ratios in the indicated strains shown in (B).Kruskal–Wallis nonparametric statistical test (****p < 0.0001). Whiskers represent mean ± 1.5 × SD (N = 3, n > 150 cells/strain). (E) Graph depicting the raw fluorescence intensity of Gin4-GFP at the large bud in the indicated strains shown in (B).Kruskal–Wallis nonparametric statistical test (***p < 0.001, ****p < 0.0001). Whiskers represent mean ± 1.5 × SD (N = 3, wild-type: n = 125, elm1Δ: n = 125, elm1Δ-Hsl1-GBP: n = 118 cells). (F) Representative images showing the localization of Gin4FL-GFP and Gin4KD-GFP constructs expressed under the pTEF promoter and their tethering to the bud neck with Hsl1-GBP in elm1Δ gin4Δ double-deletion strains. Scale bar: 5 µm. (G) Stacked column graph showing the percentage of cells exhibiting round and elongated/clumped morphologies in the indicated strains shown in (F).One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean. p-values correspond to the population of cells exhibiting round morphology (N = 3, elm1Δ gin4Δ: n = 339, elm1Δ gin4Δ-Empty vector: n = 327, elm1Δ gin4Δ-Gin4FL: n = 173, elm1Δ gin4Δ-Gin4KD: n = 236, elm1Δ gin4Δ-Empty vector-Hsl1-GBP: n = 356, elm1Δ gin4Δ-Gin4FL-Hsl1-GBP: n = 365, elm1Δ gin4Δ-Gin4KD-Hsl1-GBP: n = 459 cells). (H) Quantification of aspect ratios in the indicated strains shown in (F).Kruskal–Wallis nonparametric statistical test (****p < 0.0001, ns: p > 0.05).Whiskers represent mean ± 1.5 × SD (N = 3, n > 90 cells/strain). (I) Representative images showing artificial tethering of Gin4-GFP with Hsl1-ka1Δ-GBP in elm1Δ cells. Scale bar: 5 µm. (J) Stacked column graph showing the percentage of cells exhibiting round and elongated/clumped morphologies in the indicated strains shown in (I).One-way ANOVA with Tukey’s multiple comparison test (****p < 0.0001, ns: p > 0.05).Error bars indicate + SD from the mean. p-values correspond to the population of cells exhibiting round morphology (N = 3, wild-type: n = 674, Hsl1-ka1Δ-GBP: n = 419, elm1Δ: n = 572, elm1Δ Hsl1-ka1Δ-GBP: n = 337 cells). (K) Quantification of aspect ratios in the indicated strains shown in (I).Kruskal–Wallis nonparametric statistical test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 150 cells/strain).

Figure 4—source data 1

Numerical data underlying the quantitative analyses represented in Figure 4.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
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Targeted localization of Gin4-GFP to the bud neck in elm1Δ cells via different bud neck proteins.

(A) Representative montages showing the recruitment of Gin4-GFP during bud emergence (t0) in elm1Δ background. Scale bar: 5 µm. (B) Representative montages of the bud neck depicting the localization of Gin4-GFP in elm1Δ cells during cytokinesis (t0 = spindle breakpoint), DC* = differential contrast. (C) Graph showing the raw fluorescence intensity of Gin4-GFP in the indicated strains shown in (B) (wild-type: n = 29 and elm1Δ: n = 31 cells). (D) Plot of normalized fluorescence signal intensity of Gin4-GFP in the indicated strains shown in (B) (t0 = spindle breakpoint). (E) Representative images showing artificial tethering of Gin4-GFP via Shs1-GBP, Bud4-GBP and Bni5-GBP in elm1Δ cells. Scale bar: 5 µm. (F) Bar graph showing the percentage of cells exhibiting round morphology in the indicated strains shown in (E).One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean (N = 3, wild-type: n = 407, elm1Δ: n = 508, elm1Δ-Shs1-GBP: n = 488, elm1Δ-Bud4-GBP: n = 471, and elm1Δ-Bni5-GBP: n = 448 cells). (G) Quantification of the aspect ratios in the indicated strains shown in (E).Kruskal–Wallis nonparametric statistical test (*p < 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 155 cells/strain).

Figure 4—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 4—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig4-figsupp1-data1-v1.xlsx
Figure 5
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Redirecting Gin4 to the bud neck via Hsl1 in elm1Δ cells rescues septin organization and AMR dynamics.

(A) Representative time-lapse montages showing the rescue of mislocalization of both Gin4-GFP and Cdc3-mCherry upon artificial tethering with Hsl1-GBP in elm1Δ cells. Scale bar: 5 µm. (B) Stacked column graph showing the percentage of cells exhibiting normal septin localization or septin mislocalization to bud cortex during the bud emergence in the indicated strains shown in (A) (wild-type: n = 39, Hsl1-GBP: n = 39, elm1Δ: n = 39, elm1Δ-Hsl1-GBP: n = 42 cells). (C) Quantification of Gin4-GFP localization rescue in the indicated strains shown in (A) (wild-type: n = 39, Hsl1-GBP: n = 39, elm1Δ: n = 39, elm1Δ-Hsl1-GBP: n = 42 cells). (D) Representative montages of the bud neck showing the constriction kinetics of Myo1-ymScarletI in elm1Δ strains in which Gin4-GFP is artificially tethered to the bud neck via Hsl1-GBP. (E) Quantitative analysis for normal localization and mislocalization of Myo1-ymScarletI in the indicated strains shown in (D) (wild-type: n = 41, Hsl1-GBP: n = 37, elm1Δ: n = 39, elm1Δ-Hsl1-GBP: n = 44 cells). (F) Graph showing the constriction dynamics of Myo1-ymScarletI in the indicated strains shown in (D). Disappearance of Gin4 from the bud neck is used to normalize the initial time point for Myo1 constriction. Kruskal–Wallis nonparametric statistical test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (wild-type: n = 39, Hsl1-GBP: n = 30, elm1Δ: n = 32, elm1Δ-Hsl1-GBP: n = 39 cells).

Figure 5—source data 1

Numerical data underlying the quantitative analyses represented in Figure 5.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig5-data1-v1.xlsx
Figure 6 with 1 supplement
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Gin4 tethering to the bud neck via Hsl1 bypasses the requirement for Elm1 independently of the morphogenetic checkpoint kinase Swe1.

(A) Time-lapse montages representing the dynamics of Cdc3-mCherry in both elm1Δ and elm1Δ swe1Δ strains upon artificial tethering of Gin4-GFP to bud neck with Hsl1-GBP (t0 = bud emergence). Scale bar: 5 µm. (B) Stacked column graph showing the percentage of cells exhibiting normal localization and mislocalization of Cdc3-mCherry in the indicated strains shown in (A) (wild-type: n = 55, swe1Δ: n = 49, elm1Δ: n = 58, elm1Δ swe1Δ: n = 51, elm1Δ-Gin4-GFP: n = 55, elm1Δ swe1Δ-Gin4-GFP: n = 55, elm1Δ-Gin4-GFP-Hsl1-GBP: n = 58, elm1Δ swe1Δ-Gin4-GFP-Hsl1-GBP: n = 52 cells). (C) Representative time-series montages showing the localization of Gin4-GFP in both elm1Δ and elm1Δ swe1Δ strains upon artificial tethering of Gin4-GFP to the bud neck via Hsl1-GBP (t0 = bud emergence). Scale bar: 5 µm. (D) Quantification graphs depicting the normal localization and mislocalization of Gin4-GFP at the bud cortex during bud emergence in the indicated strains shown in (C) (wild-type: n = 57, swe1Δ: n = 52, Hsl1-GBP: n = 54, Hsl1-GBP swe1Δ: n = 51, elm1Δ-Gin4-GFP: n = 55, elm1Δ swe1Δ-Gin4-GFP: n = 55, elm1Δ-Gin4-GFP-Hsl1-GBP: n = 58, elm1Δ swe1Δ-Gin4-GFP-Hsl1-GBP: n = 52 cells).

Figure 6—source data 1

Numerical data underlying the quantitative analyses represented in Figure 6.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
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Absence of Swe1 rescues morphological defects observed in elm1Δ cells.

(A) Brightfield images representing cellular morphology in the elm1Δ and elm1Δ swe1Δ cells in which Gin4-GFP is artificially tethered to bud neck via Hsl1-GBP. Scale bar: 5 µm. (B) Bar graph representing the percentage of cells exhibiting round morphology in the indicated strains shown in (A).One-way ANOVA with Tukey’s multiple-comparison test (**p < 0.01, ****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean (N = 3, wild-type: n = 706, swe1Δ: n = 540, elm1Δ: n = 436, elm1Δ swe1Δ: n = 638, elm1Δ-Gin4-GFP: n = 450, elm1Δ swe1Δ-Gin4-GFP: n = 465, elm1Δ-Gin4-GFP-Hsl1-GBP: n = 623, elm1Δ swe1Δ-Gin4-GFP-Hsl1-GBP: n = 604 cells). (C) Quantification of the aspect ratios in the indicated strains shown in (A). Kruskal–Wallis nonparametric statistical test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 170 cells/strain).

Figure 6—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 6—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig6-figsupp1-data1-v1.xlsx
Figure 7 with 2 supplements
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Restoring Hsl1-GFP localization to the bud neck via septins or its related kinases can bypass the requirement of Elm1.

(A) Representative images showing artificial tethering of Hsl1-GFP to the bud neck via Shs1-GBP, Bud4-GBP, Kcc4-GBP, Bni5-GBP, and Gin4-GBP in elm1Δ cells. Scale bar: 5 µm. (B) Bar graph showing the percentage of cells exhibiting round morphology in the indicated strains shown in (A).One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean (N = 3, wild-type: n = 724, elm1Δ: n = 458, elm1Δ-Shs1-GBP: n = 726, elm1Δ-Bud4-GBP: n = 717, elm1Δ-Kcc4-GBP: n = 862, elm1Δ-Bni5-GBP: n = 358, and elm1Δ-Gin4-GBP: n = 754 cells). (C) Quantification of aspect ratios in the indicated strains shown in (A).Kruskal–Wallis nonparametric statistical test (***p < 0.001, ****p < 0.0001). Whiskers represent mean ± 1.5 × SD (N = 3, n > 135 cells/strain). (D) Representative montages of the bud neck depicting the localization of Hsl1-GFP in elm1Δ and gin4Δ cells during cytokinesis. DC* = differential contrast. (E) Plot of raw fluorescence intensity of Hsl1-GFP in the indicated strains shown in (D) (t0 = spindle breakpoint) (wild-type: n = 29, elm1Δ: n = 24, gin4Δ: n = 38 cells). (F) Plot of normalized fluorescence intensity of Hsl1-GFP in the indicated strains shown in (D) (t0 = spindle breakpoint).

Figure 7—source data 1

Numerical data underlying the quantitative analyses represented in Figure 7.

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Figure 7—figure supplement 1
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Redirecting Kcc4-GFP localization to bud neck via Hsl1-GBP rescues defects associated with elm1Δ cells.

(A) Representative images showing artificial tethering of Kcc4-GFP to the bud neck via Shs1-GBP, Bud4-GBP, Bni5-GBP, Gin4-GBP, and Hsl1-GBP in elm1Δ cells. Scale bar: 5 µm. (B) Bar graph showing the percentage of cells exhibiting round morphology in the indicated strains shown in (A). One-way ANOVA with Tukey’s multiple-comparison test (*p < 0.05, **p < 0.01, ****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean (N = 3, wild-type: n = 724, elm1Δ: n = 491, elm1Δ-Shs1-GBP: n = 587, elm1Δ-Bud4-GBP: n = 663, elm1Δ-Bni5-GBP: n = 569, elm1Δ-Gin4-GBP: n = 483 and elm1Δ-Hsl1-GBP: n = 708 cells). (C) Quantification of aspect ratios in the indicated strains shown in (A).Kruskal–Wallis nonparametric statistical test (***p < 0.001, ****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 155 cells/strain).

Figure 7—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 7—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig7-figsupp1-data1-v1.xlsx
Figure 7—figure supplement 2
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Kcc4 is recruited to the bud neck in an Elm1-dependent and Gin4-independent manner.

(A) Representative montages showing the recruitment of Hsl1-GFP during bud emergence (t0) in elm1Δ and gin4Δ backgrounds. DC* = differential contrast. Scale bar: 5 µm. (B) Representative montages showing the recruitment of Kcc4-GFP during bud emergence (t0) in elm1Δ and gin4Δ backgrounds. DC* = differential contrast. Scale bar: 5 µm. (C) Representative montages of the bud neck depicting the localization of Kcc4-GFP in elm1Δ and gin4Δ cells during cytokinesis. (D) Plot of raw fluorescence intensity of Kcc4-GFP in the indicated strains shown in (C) (t0 = spindle breakpoint) (wild-type: n = 30, elm1Δ: n = 26, and gin4Δ: n = 30 cells). (E) Plot of normalized fluorescence intensity of Kcc4-GFP in indicated strains shown in (C) (t0 = spindle breakpoint).

Figure 7—figure supplement 2—source data 1

Numerical data underlying the quantitative analyses represented in Figure 7—figure supplement 2.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig7-figsupp2-data1-v1.xlsx
Figure 8 with 2 supplements
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A non-canonical role for Hsl1-Kinase in regulating septin organization and AMR dynamics.

(A) Representative images showing Cdc3-mCherry mislocalization during bud emergence in gin4Δ and gin4Δ hsl1Δ cells in which Elm1-GFP is artificially tethered to bud neck via Shs1-GBP. White arrows indicate mislocalized Cdc3-mCherry in the represented strains. Scale bar: 5 µm. (B) Representative time-lapse montages of the bud neck showing the constriction profile of Inn1-3xmCherry in gin4Δ and gin4Δ hsl1Δ cells in which Elm1-GFP is artificially tethered to bud neck via Shs1-GBP. (C) Bar graph showing the percentage of cells exhibiting round and elongated morphologies in the indicated strains shown in (A).One-way ANOVA with Tukey’s multiple-comparison test (**p < 0.01, ***p < 0.001, ns: p > 0.05). Error bars indicate + SD from the mean. p-values correspond to population of cells exhibiting round morphology (N = 3, gin4Δ: n = 689, gin4Δ-Shs1-GBP: n = 639, gin4Δ hsl1Δ: n = 492, gin4Δ hsl1Δ-Shs1-GBP: n = 590 cells). (D) Quantification of aspect ratios in the indicated strains shown in (A). Kruskal–Wallis nonparametric statistical test (****:p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 165 cells/strain). (E) Stacked column graph representing the percentage of cells exhibiting normal septin localization and septin mislocalization in the indicated strains of (A) (gin4Δ: n = 44, gin4Δ-Shs1-GBP: n = 41, gin4Δ hsl1Δ: n = 38, gin4Δ hsl1Δ-Shs1-GBP: n = 44 cells). (F) Quantification of normal localization and mislocalization of Elm1-GFP in the indicated strains shown in (A) (gin4Δ-Shs1-GBP: n = 37, gin4Δ hsl1Δ-Shs1-GBP: n = 44 cells) (G) Quantification of the residence time of Inn1-3xmcherry during cytokinesis in the indicated strains shown in (B). Kruskal–Wallis nonparametric statistical test (*p < 0.05, **p < 0.01, ****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 2, n > 60 cells/strain).

Figure 8—source data 1

Numerical data underlying the quantitative analyses represented in Figure 8.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig8-data1-v1.xlsx
Figure 8—figure supplement 1
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Hsl1 kinase becomes essential downstream of Elm1 to regulate cytokinesis in gin4Δ cells.

(A) Representative montages showing the recruitment of Elm1-GFP during bud emergence (t0) in gin4Δ background. Scale bar: 5 µm. (B) Representative montages of the bud neck depicting the localization of Elm1-GFP during cytokinesis in gin4Δ cells. DC* = differential contrast. (C) Plot representing the raw fluorescence intensity of Elm1-GFP in the indicated strains shown in (B) (t0 = spindle breakpoint) (wild-type: n = 26 and gin4Δ: n = 32 cells). (D) Plot of normalized fluorescence intensity of Elm1-GFP in indicated strains of (B) (t0 = spindle breakpoint). (E) Representative images showing artificial tethering of Elm1-GFP via Shs1-GBP, Bud4-GBP, and Hsl1-GBP in gin4Δ cells. Scale bar: 5 µm. (F) Bar graph showing the percentage of cells exhibiting round morphology in the indicated strains shown in (E).One-way ANOVA with Tukey’s multiple-comparison test (*p < 0.05, **p < 0.01, ***p < 0.001, ns: p > 0.05). Error bars indicate + SD from the mean (N = 3, wild-type: n = 389, gin4Δ: n = 415, gin4Δ-Shs1-GBP: n = 800, gin4Δ-Bud4-GBP: n = 688 and gin4Δ-Hsl1-GBP: n = 886 cells). (G) Quantification of aspect ratios in the indicated strains shown in (E).Kruskal–Wallis nonparametric statistical test (**p < 0.01, ****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 165 cells/strain). (H) Representative images showing artificial tethering of Elm1-GFP with Hsl1-Δka1-GBP in gin4Δ cells. Scale bar: 5 µm. (I) Bar graph showing the percentage of cells exhibiting round and elongated/clumped morphologies in the indicated strains shown in (H).One-way ANOVA with Tukey’s multiple-comparison test (****p < 0.0001, ns: p > 0.05). Error bars indicate + SD from the mean. p-values correspond to the population of cells with round morphology (N = 3, wild-type: n = 765, Hsl1-GBP: n = 663, gin4Δ: n = 602, and gin4Δ-Hsl1-GBP: n = 609 cells). (J) Quantification of aspect ratios in the indicated strains shown in (H).Kruskal–Wallis nonparametric statistical test (****p < 0.0001, ns: p > 0.05). Whiskers represent mean ± 1.5 × SD (N = 3, n > 165 cells/strain).

Figure 8—figure supplement 1—source data 1

Numerical data underlying the quantitative analyses represented in Figure 8—figure supplement 1.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig8-figsupp1-data1-v1.xlsx
Figure 8—figure supplement 2
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Absence of Hsl1 exacerbates the mislocalization of Myo1 to bud cortex in gin4Δ cells.

(A) Representative images showing mislocalization of Myo1-3xmCherry to the bud cortex in gin4Δ and gin4Δ hsl1Δ cells in which Elm1-GFP is artificially tethered to the bud neck via Shs1-GBP. White arrows indicate mislocalized Myo1-3xmCherry in the represented strains. Scale bar: 5 µm. (B) Stacked column graph showing the percentage of cells exhibiting normal localization and mislocalization of Myo1-3xmCherry during bud emergence in the indicated strains shown in (A) (gin4Δ: n = 39, gin4Δ-Shs1-GBP: n = 36, gin4Δ hsl1Δ: n = 38, and gin4Δ hsl1Δ-Shs1-GBP: n = 39 cells).

Figure 8—figure supplement 2—source data 1

Numerical data underlying the quantitative analyses represented in Figure 8—figure supplement 2.

https://cdn.elifesciences.org/articles/106366/elife-106366-fig8-figsupp2-data1-v1.xlsx
Figure 9
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Representative model for the role of Hsl1 kinase in septin organization and AMR constriction downstream of Gin4 and Elm1.

Created with BioRender.com.

Author response image 1
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Plot showing spatiotemporal kinetics of Cdc3-mCherry in strains expressing either Elm1-GFP, or Gin4-GFP, or Hsl1-GFP, or Kcc4-GFP.
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Artificial tethering of Gin4-GFP to the bud neck via Hsl1-GBP-ymScarletI rescues cellular morphology in elm1Δ cells.

(A) Representative images showing artificial tethering of Gin4-GFP via Shs1-GBP-ymScarletI, Hsl1-GBP-ymScarletI, Bud4-GBPymScarletI and Bni5-GBP-ymScarletI in elm1Δ cells. DC*=Differential contrast. Scale bar5µm. (B) Bar graph representing the percentage of cells exhibiting round morphology in the indicated strains shown in (A), one-way ANOVA Tukey’s multiple-comparison test (**: p<0.01, ****: p<0.0001, ns: p>0.05), (N=3, wildtype: n=397, elm1Δ: n=408, elm1Δ-Shs1-GBPymScarletI: n=467, elm1Δ-Hsl1-GBP-ymScarletI: n=462, elm1Δ-Bud4-GBP-ymScarletI: n=401 and elm1Δ-Bni5-GBP-ymScarletI: n=317 cells). (C) Quantification of aspect ratios in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (***: p<0.001, ****: p<0.0001, ns: p>0.05), (N=3, n>170 cells/strain). (D) Graph depicting the raw fluorescence intensity of Gin4-GFP at the large bud in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (***: p<0.001, ****: p<0.0001, ns: p>0.05), (N=3, wildtype: n=166, elm1Δ: n=177, elm1Δ-Shs1-GBP-YmScarletI: n=176, elm1Δ-Hsl1-GBPymScarletI: n=188, elm1Δ-Bud4-GBP-ymScarletI: n=185 and elm1Δ-Bni5-GBP-ymScarletI: n=151 cells).

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Artificial tethering of Hsl1-GFP to the bud neck via septins or Nim1-related kinases rescues cellular morphology in elm1Δ cells.

(A) Representative images showing the relocalization of Hsl1-GFP to the bud neck in elm1Δ cells via Shs1-GBP-ymScarletI and Gin4GBP-ymScarletI. Scale bar-5µm. (B) Bar graph representing the percentage of cells exhibiting round morphology in the indicated strains shown in (A), one-way ANOVA Tukey’s multiple comparison test (****: p<0.0001, ns: p>0.05), (N=3, wildtype: n=508, elm1Δ: n=418, elm1ΔShs1-GBP-ymScarletI: n=535 and elm1Δ-Gin4-GBP-ymScarletI: n=482 cells). (C) Quantification of the aspect ratios in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (*: p<0.05, ****: p<0.0001), (N=3, n>165 cells/strain). (D) Quantification of raw fluorescence intensity of Hsl1-GFP at the large bud in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (***: p<0.001, ****: p<0.0001), (N=3, wildtype: n=156, elm1Δ: n=166, elm1Δ-Shs1-GBP-ymScarletI: n=169 and elm1Δ-Gin4-GBP-ymScarletI: n=165 cells).

Author response image 5
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Targeted localization of Kcc4-GFP to the bud neck via Hsl1-GBP-ymScarletI rescues cellular morphology in elm1Δ cells.

(A) Representative images showing artificial tethering of Kcc4-GFP to the bud neck in elm1Δ cells via Shs1-GBP-ymScarletI, Hsl1-GBPymScarletI and Gin4-GBP-ymScarletI. Scale bar-5µm. (B) Quantitative analysis representing the percentage of cells exhibiting round morphology in the indicated strains shown in (A), oneway ANOVA Tukey’s multiple-comparison test (****: p<0.0001, ns: p>0.05), (N=3, wildtype: n=443, elm1Δ: n=489, elm1Δ-Shs1-GBP-ymScarletI: n=312, elm1Δ-Hsl1-GBP-ymScarletI: n=563 and elm1Δ-Gin4-GBP-ymScarletI: n=337 cells). (C) Quantification of the aspect ratios in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (****: p<0.0001, ns: p>0.05), (N=3, n>165 cells/strain). (D) Quantification for the raw fluorescence intensity of Kcc4-GFP at the large bud in the indicated strains shown in (A), Kruskal-Wallis nonparametric statistical test (**: p<0.01, ***: p<0.001, ****: p<0.0001), (N=3, wildtype: n=163, elm1Δ: n=168 elm1Δ-Shs1-GBP-ymScarletI: n=161, elm1Δ-Hsl1-GBP-ymScarletI: n=172 and elm1Δ-Gin4-GBP-ymScarletI: n=150 cells).

Tables

Table 1
Table summarizing the rescue of elongated or clumped morphology upon redirecting the Nim1-related kinases to the bud neck in elm1Δ cells and Elm1-GFP in gin4Δ cells using GFP-GBP artificial tethering strategy.
Protein tagged with GFPProtein tagged with GBPGenetic backgroundNumber of cells exhibiting round morphologyNumber of cells exhibiting elongated/clumped morphologyTotal number of cellsPercentage of cellular population exhibiting round morphologyRescue of elongated or clumped morphology
Gin4Shs1elm1Δ244644884.91No
Gin4Bud4elm1Δ204514714.24No
Gin4Hsl1elm1Δ25514540063.75Yes
Gin4Hsl1-ka1Δelm1Δ63313371.78No
Gin4Bni5elm1Δ174314483.79No
Gin4-Full lengthHsl1elm1Δ gin4Δ2828336577.26Yes
Gin4-Kinase deadHsl1elm1Δ gin4Δ30115845965.57Yes
Hsl1Shs1elm1Δ26446272636.36Yes
Hsl1Bud4elm1Δ11060771715.34No
Hsl1Bni5elm1Δ43543581.11No
Hsl1Gin4elm1Δ52123375469.09Yes
Hsl1Kcc4elm1Δ49836486257.77Yes
Kcc4Shs1elm1Δ215665873.57No
Kcc4Bud4elm1Δ606036639.04No
Kcc4Hsl1elm1Δ51619270872.88Yes
Kcc4Bni5elm1Δ65635691.05No
Kcc4Gin4elm1Δ11936448324.63No
Elm1Bud4gin4Δ61627088669.52Yes
Elm1Hsl1gin4Δ39928968857.99Yes
Elm1Hsl1-ka1Δgin4Δ13947060922.82No
Elm1Shs1gin4Δ49930180062.37Yes

  1. Bold text indicates conditions in which artificial tethering resulted in rescue of the elongated or clumped cellular morphology, as indicated in the final column.

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain backgroundEscherichia coli-DH5aNew England BiolabsCatalog # C2987I
Strain, strain backgroundEscherichia coli-Top10Thermo Fisher ScientificCatalog number: C404003
Strain, strain backgroundEscherichia coli-BL21(DE3)New England BiolabsCatalog #
C2527
Strain, strain backgroundSaccharomyces cerevisiae S288C-background strainsThis paperYeast strains used in this study are listed in Supplementary file 1. Both previously reported and newly generated strains, together with their respective references, are indicated therein. All strains are available from the corresponding author upon request.
Chemical compound, drugG418-DisulphateSisco Research Laboratories Pvt. Ltd.Cat#:58327
Chemical compound, drugHygromycin BSisco Research Laboratories Pvt. Ltd.Cat#:67317
Chemical compound, drugNourseothricinJena BioscienceCat#:AB-102
Chemical compound, drugAmpicillinG-BiosciencesCat#AB1005Lot#201512
Chemical compound, drugKanamycinG-BiosciencesCat#AB1025Lot#230201
Commercial assay or kitNEBuilder HiFi DNA Assembly Master MixNew England BiolabsCat#:E262L
Commercial assay or kitGET Plasmid MiniprepG-BiosciencesCat#:786-361
Commercial assay or kitQIAquick Gel Extraction KitQIAGENCat#:28704
Commercial assay or kitGeneJET PCR Purification KitThermo Fisher ScientificCat#:K0701
Sequence-based reagentPrimers used in this studyThis paperOligonucleotide sequences used in this study are provided in Supplementary file 2.
Recombinant DNA reagent/ PlasmidsPlasmids used in this studyThis paperSupplementary file 3 contains a list of all plasmids used in this study, including newly generated constructs and plasmids described previously, along with their respective references. These plasmids will be made available by the corresponding author upon request.
Software, algorithmFijiFijiRRID:SCR_002285https://imagej.net/software/fiji/
Software, algorithmOriginPro Pro (2015, Sr2, 69.2.272)OriginLab CorporationRRID:SCR_014212https://www.originlab.com/
Software, algorithmCellSens Dimension 3.1OlympusRRID:SCR_014551https://www.olympus-lifescience.com/en/software/cellsens/
Software, algorithmGraphPad Prism 6.04GraphPad SoftwareRRID:SCR_002798https://www.graphpad.com/
Software, algorithmAndor FusionOxford InstrumentsRRID:SCR_023609https://andor.oxinst.com/downloads/view/fusion-release-2.3
Software, algorithmInkscapeInkscape ProjectRRID:SCR_014479Used for figure preparation
Software, algorithmBioRenderBioRender.comRRID:SCR_018361Used for figure preparation (Figures 2I, 4A, and 9)
Software, algorithmAdobe IllustratorAdobeRRID:SCR_010279Used for figure preparation (Figure 2F, G, Figure 2—figure supplement 1H)
Antibodyanti-His (H-3), mouse monoclonal IgG1Santa Cruz BiotechnologyCat#:sc-8036, RRID:AB_627727WB; (1:2000); Lot#:C0421
Antibodyanti-GST (B-14), mouse monoclonal IgG1Santa Cruz BiotechnologyCat#:sc-138, RRID:AB_627677WB; (1:2000); Lot#:K1020
Antibodyanti-mouse IgG-HRPCell Signaling TechnologyCat#:7076S, RRID:AB_330924WB; (1:3000): Lot#:36
OtherConcanavalin A, type VISigma-AldrichCat#:C2010
OtherConfocal dish (35 mm)ibidi GmbHCat#:81218-200
OtherConfocal dish (35 mm)CellvisCat#:D35C4-20-1.5-N
OtherNi-NTA agarose resinG-BiosciencesCat#:786-940Affinity resin used for purification of 6His-bdSUMO-Gin4KA1
OtherGlutathione Sepharose 4B resinGE HealthcareCat#:GE17-0756-01Affinity resin used for purification/immobilization of GST-Hof1N-ter
OtherPoly-Prep chromatography columnsBio-Rad LaboratoriesCat#:731-1550Used for affinity purification
OtherPD Midi Trap G-10 columnsGE HealthcareCat#:GE28-9180-11Used for protein buffer exchange
Author response table 1
Summary table showing rescue of elongated morphology in elm1Δ cells upon forced recruitment of Nim1-related kinases via septins or its related kinases tagged with GBPymScarletI.
Protein tagged with GFPProtein tagged with GBPGenetic backgroundNumber of cells exhibiting round morphologyNumber of cells exhibiting elongated/clumped morphologyTotal number of cellsPercentage of cellular population exhibiting round morphologyRescue of elongated or clumped morphology
Gln4Shs1elm1△374304677.92No
GIn4Bud4elm1△5234940112.96No
Gln4Hsl1elm1 △3649846278.78Yes
Gln4BnI5elm1△03173170No
Hsl1Shs16/m1 △33320253562.24Yes
Hsl1Gln4elm1 △25822448253.52Yes
Kcc4Shs1elm1 △242883127.69No
Kcc4Hsl1eim1 △40216156371.40Yes
Kco4Gln4eimf 1Delta113263373.26No

Additional files

Supplementary file 1

List of yeast strains used in this study.

https://cdn.elifesciences.org/articles/106366/elife-106366-supp1-v1.docx
Supplementary file 2

List of oligonucleotides used in this study.

https://cdn.elifesciences.org/articles/106366/elife-106366-supp2-v1.docx
Supplementary file 3

List of plasmids used in this study.

https://cdn.elifesciences.org/articles/106366/elife-106366-supp3-v1.docx
MDAR checklist
https://cdn.elifesciences.org/articles/106366/elife-106366-mdarchecklist1-v1.docx

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  1. Bindu Bhojappa
  2. Anubhav Dhar
  3. VT Bagyashree
  4. Jayanti Kumari
  5. Freya Cardozo
  6. Vaseef Rizvi
  7. Deepthi Guturu
  8. Saravanan Palani
(2026)
Nim1-related kinases regulate septin organization and cytokinesis by modulating Hof1 at the cell division site
eLife 14:RP106366.
https://doi.org/10.7554/eLife.106366.3