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Effects of rok1 gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms

  • Authors:
    • Jiayi He
    • Mengnan Liu
    • Jiani Xu
    • Xiang Ding
    • Yiling Hou
  • View Affiliations / Copyright

    Affiliations: Key Laboratory of Southwest China Wildlife Resources Conservation, Ministry of Education, College of Life Science, China West Normal University, Nanchong, Sichuan 637009, P.R. China, Nanchong Key Laboratory of Eco‑Environmental Protection and Pollution Control in Jialing River Basin, College of Environmental Science and Engineering, China West Normal University, Nanchong, Sichuan 637009, P.R. China
    Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 151
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    Published online on: April 2, 2026
       https://doi.org/10.3892/etm.2026.13145
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Abstract

The rok1 gene encodes the ATP‑dependent RNA helicase Rok1, which is involved in regulating the maturation of small subunit ribosomal RNA and thus ribosome biogenesis. However, the regulation of cellular mitotic dynamics by the rok1 gene deletion is currently unclear. In the present study, fluorescent protein labeling and live cell imaging techniques were used to investigate the effects of rok1 deletion on the dynamics of microtubules, actin and kinetochores during mitosis at 25 and 37˚C, and RNA‑sequencing and bioinformatics analyses were used to reveal the key genes. Analysis of the live cell imaging results revealed that, in mitosis, the initiation length and contraction length of actin rings were both shortened and the contraction rate was decreased at 25 and 37˚C. The separation process of kinetochores was inhibited at 25 and 37˚C, and the inhibition was more severe at the higher temperature of 37˚C. Analysis of RNA sequencing results showed that upregulation of myo51 and blt1 resulted in delayed actin ring assembly and slowed actin ring contraction in the rok1Δ strain. In addition, psm1 and psc3 were upregulated and are key genes affecting the ability of kinetochores to move on the spindle and the cohesion of sister chromatids. The present study revealed that the Rok1 protein not only influences the actin polymerization process, participate in the regulation of actin ring assembly and contraction, and cytoplasmic division, but also affects the migration ability of kinetochores on the spindle and participate in the regulation of the formation and maintenance of cohesion between sister chromatids, which provides a certain scientific basis for further exploring the function of the Rok1 protein in cell division.
View Figures

Figure 1

Location, protein structure and
protein conserved domain of rok1. (A) Location of
rok1. (B) Structure of the Rok1 protein. (C) Conserved
domains of the Rok1 protein.

Figure 2

Effects of rok1 deletion on
cell growth and ascospores. (A) Growth rate analysis of wt and
rok1Δ strains at 25˚C. (B) Growth rate analysis of wt and
rok1Δ strains at 37˚C. (C) Spore morphology of wt and
rok1Δ strains at 25˚C. (D) Number of spores in wt and
rok1Δ strains. *P<0.05 and
**P<0.01; n=1,200. wt, wild-type; OD, optical
density.

Figure 3

Effects of rok1 deletion on
spindle dynamics during mitosis in fission yeast. (A) Images of
spindles during mitosis in wt and rok1Δ cells at 25˚C and
37˚. (B) Spindle length changes of wt and rok1Δ cells during
mitosis at 25˚C. (C) Spindle length changes of wt and rok1Δ
cells during mitosis at 37˚C. (D) Spindle elongation lengths of wt
and rok1Δ cells at different phases. (E) Spindle elongation
time of wt and rok1Δ cells at different phases. (F) Spindle
elongation rates of wt and rok1Δ cells at different phases.
*P<0.05 and **P<0.01; n=15. GFP, green
fluorescent protein; RFP, red fluorescent protein; wt,
wild-type.

Figure 4

Effects of rok1 deletion on
actin dynamics during mitosis. (A) Images of the actin ring during
mitosis in wt and rok1Δ cells at 25˚C and 37˚C. (B) Actin
ring length changes of wild-type and rok1Δ cells during
mitosis at 25˚C. (C) Actin ring length changes of wild-type and
rok1Δ cells during mitosis at 37˚C. (D) The length of the
actin ring of wild-type and rok1Δ cells. (E) The time of the
actin ring formation and contraction of wild-type and rok1Δ
cells at different phases. (F) The rate of the actin ring
contraction of wild-type and rok1Δ cells at different
phases. *P<0.05 and **P<0.01; n=15.
GFP, green fluorescent protein; wt, wild-type.

Figure 5

Effects of rok1 deletion on
kinetochore dynamics during mitosis in fission yeast. (A) Images of
kinetochore during mitosis in wild-type and rok1Δ cells at
25˚C and 37˚C, white arrows indicated the position of kinetochores.
(B) Kinetochore separation distance changes of wild-type and
rok1Δ cells during mitosis at 25˚C. (C) Kinetochore
separation distance changes of wild-type and rok1Δ cells
during mitosis at 37˚C. (D) Time from the separation of the
kinetochore to localize at the ends of the spindle in the wild-type
and rok1Δ cells. (E) Kinetochore separation distance of
wild-type and rok1Δ cells. (F) Kinetochore separation time
of wild-type and rok1Δ cells. (G) Kinetochore separation
rates of wild-type and rok1Δ cells. *P<0.05
and **P<0.01; n=15. GFP, green fluorescent protein;
wt, wild-type.

Figure 6

Effects of rok1 deletion on
centrosome dynamics during mitosis in fission yeast. (A) Images of
centrosome during mitosis in wt and rok1Δ cells at 25˚C and
37˚C, white arrows indicated the position of centrosome. (B)
Centrosome separation distance changes of wt and rok1Δ cells
during mitosis at 25˚C. (C) Centrosome separation distance changes
of wt and rok1Δ cells during mitosis at 37˚C. (D) Centrosome
separation distance in the wt and rok1Δ cells at spindle
breakage. (E) Centrosome separation time of wt and rok1Δ
cells at different phases. (F) Centrosome separation rates of wt
and rok1Δ cells at different phases. *P<0.05
and **P<0.01; n=15. GFP, green fluorescent protein;
wt, wild-type.

Figure 7

Analysis of highly expressed genes
and differentially expressed genes. (A) Differential analysis of
highly expressed genes in the wt and rok1Δ strains at 25˚C.
(B) Differential analysis of highly expressed genes in the wt and
rok1Δ strains at 37˚C. (C) Volcano plot of differently
expressed genes in the wt and rok1Δ strains at 25˚C. (D)
Volcano plot of differently expressed genes in the wt and
rok1Δ strains at 37˚C. *P<0.05 and
**P<0.01. n=3. FPKM, fragments per kilobase million;
wt, wild-type.

Figure 8

RT-qPCR verification of key
differentially expressed genes. (A) Relative mRNA expression of
psc3 and psm1 determined by RT-qPCR at 25˚C. (B)
Relative mRNA expression of myo51 and blt1 determined
by RT-qPCR at 37˚C. *P<0.05 and
**P<0.01. RT-qPCR, reverse transcription quantitative
PCR; wt, wild-type.

Figure 9

GO enrichment results of
differentially expressed genes in the wild-type and rok1Δ
strains at 25˚C. (A) GO enrichment results for upregulated
differential genes at 25˚C. (B) GO enrichment results for
downregulated differential genes at 25˚C. GO, Gene Ontology; BP,
biological process; CC, cellular component; MF, molecular
function.

Figure 10

GO enrichment results of
differentially expressed genes in the wild-type and rok1Δ
strains at 37˚C. (A) GO enrichment results for upregulated
differential genes at 37˚C. (B) GO enrichment results for
downregulated differential genes at 37˚C. GO, Gene Ontology; BP,
biological process; CC, cellular component; MF, molecular
function.

Figure 11

KEGG enrichment results of
differentially expressed genes in the wild-type and rok1Δ
strains. (A) KEGG enrichment results for upregulated and
downregulated differential genes at 25˚C. (B) KEGG enrichment
results for upregulated and downregulated differential genes at
37˚C. KEGG, Kyoto Encyclopedia of Genes and Genomes.
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Copy and paste a formatted citation
Spandidos Publications style
He J, Liu M, Xu J, Ding X and Hou Y: Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms. Exp Ther Med 31: 151, 2026.
APA
He, J., Liu, M., Xu, J., Ding, X., & Hou, Y. (2026). Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms. Experimental and Therapeutic Medicine, 31, 151. https://doi.org/10.3892/etm.2026.13145
MLA
He, J., Liu, M., Xu, J., Ding, X., Hou, Y."Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms". Experimental and Therapeutic Medicine 31.6 (2026): 151.
Chicago
He, J., Liu, M., Xu, J., Ding, X., Hou, Y."Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms". Experimental and Therapeutic Medicine 31, no. 6 (2026): 151. https://doi.org/10.3892/etm.2026.13145
Copy and paste a formatted citation
x
Spandidos Publications style
He J, Liu M, Xu J, Ding X and Hou Y: Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms. Exp Ther Med 31: 151, 2026.
APA
He, J., Liu, M., Xu, J., Ding, X., & Hou, Y. (2026). Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms. Experimental and Therapeutic Medicine, 31, 151. https://doi.org/10.3892/etm.2026.13145
MLA
He, J., Liu, M., Xu, J., Ding, X., Hou, Y."Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms". Experimental and Therapeutic Medicine 31.6 (2026): 151.
Chicago
He, J., Liu, M., Xu, J., Ding, X., Hou, Y."Effects of <em>rok1</em> gene deletion on mitosis in fission yeast at appropriate and stressful temperatures and the molecular mechanisms". Experimental and Therapeutic Medicine 31, no. 6 (2026): 151. https://doi.org/10.3892/etm.2026.13145
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