Effects of <i>pmr1</i> Gene Deletion on Sexual Reproduction and Mitosis of Fission Yeast Cells and Its Molecular Mechanism

doi:10.13523/j.cb.2204077

China Biotechnology

2022, Vol. 42

Issue (12): 12-26 DOI: 10.13523/j.cb.2204077

Effects of pmr1 Gene Deletion on Sexual Reproduction and Mitosis of Fission Yeast Cells and Its Molecular Mechanism

YE Zi-yu¹,DING Xiang²,LU Yan¹,ZHOU Li-qian¹,LIU Xin-lan²,PU Di-hong¹,HOU Yi-ling^1,^**(

)

1 Key Laboratory of Southwest China Wildlife Resource Conservation (Ministry of Education), College of Life Science, China West Normal University, Nanchong 637009, China
2 College of Environmental Science and Engineering, China West Normal University, Nanchong 637009, China

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Abstract

Objective: The pmr1 gene encodes P-type calcium-transporting ATPase Pmr1, which is involved in maintaining cell wall integrity and regulating cytokinesis. In this study, fission yeast was used as a model cell to explore the effects of pmr1 deletion on the sexual reproduction and dynamics of actin rings during cell mitosis, and to reveal the key genes and metabolic pathways of abnormal cell mitosis after pmr1 deletion. Methods: The effects of pmr1 deletion on cell mitosis and sexual reproduction were detected by cell growth rate measurement, spore production observation and statistics, green fluorescent protein-labeled monitoring and living cell imaging; The wild-type and pmr1Δ strain were sequenced by RNA-Seq and analyzed by bioinformatics, and verified by qRT-PCR. Results: The pmr1 deletion could slow down cell growth, decrease cell length in mitosis, increase length of sexually reproductive ascospore, and increase formation time of actin ring. RNA-sequencing results revealed that down-regulation of mfm1, mfm2 and mat1-Mc, up-regulation of cdc1 and exo1 in the mismatch repair pathway, and down-regulation of pgi1, pfk1 and dld1 in the glycolysis/gluconeogenesis pathway are the main factor of the increased spore length in the pmr1Δ; Meanwhile, the down-regulation of tdh1 and pgk1 in the glycolysis/gluconeogenesis pathway and the down-regulation of fas1, fas2, cut6 and lcf1 in the fatty acid anabolic pathway also led to the decreased cell length in mitosis of pmr1Δ; The up-regulation of hsp9 is the key gene that affected the formation time of actin ring in pmr1Δ. qRT-PCR experiments confirmed that the expression trends of key genes after pmr1 deletion were consistent with the RNA-Seq results. Conclusion: After pmr1 deletion, the mismatch repair pathway, glycolysis/gluconeogenesis pathway and fatty acid anabolic pathway are hindered in fission yeast cells, resulting in abnormal cell and spore morphology, obstruction of actin ring formation, and slowing down of cell proliferation.

Key words： pmr1 gene Fission yeast Cell division RNA-Seq Metabolic pathway

Received: 29 April 2022 Published: 05 January 2023

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Corresponding Authors: Yi-ling HOU E-mail: starthlh@126.com

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	Zi-yu YE
	Xiang DING
	Yan LU
	Li-qian ZHOU
	Xin-lan LIU
	Di-hong PU
	Yi-ling HOU

Cite this article:

YE Zi-yu,DING Xiang,LU Yan,ZHOU Li-qian,LIU Xin-lan,PU Di-hong,HOU Yi-ling. Effects of pmr1 Gene Deletion on Sexual Reproduction and Mitosis of Fission Yeast Cells and Its Molecular Mechanism. China Biotechnology, 2022, 42(12): 12-26.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2204077 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I12/12

Fig.1 Location, protein structure and protein conserved domain of pmr1 gene (a) Location of pmr1 gene (b) Pmr1 protein structure (c) Pmr1 protein conserved domain

Table 1 Strains and genotypes in the experimental

Fig.2 Effects of pmr1 gene deletion on cell growth and ascospores at 25℃ (a) Growth rate of strain (b) Spore morphology of cell (c) Statistics of sporulation (n=150) (d) Statistics of ascospore length (n=120)

Fig.3 Effects of pmr1 gene deletion on cell length and actin ring localization (a) Five periods of dynamic changes of actin in cells (b) The initial length of cells in interphase (c) Length of cells in metaphase (d) Length of the contractile ring from the proximal cell wall (e) Length of the contractile ring from the distal cell wall (f) The ratio of the length of the contractile ring to the cell wall at both ends (n=60)

Fig.4 Effects of pmr1 gene deletion on formation time, contraction time and contraction rate of actin ring (a) Time-lapse images of actin ring dynamics (b) Kinetic curve of actin ring (c) The initial length of actin ring (d) The formation time of actin ring (e) The contraction time and contraction rate of actin ring (n=30)

Table 2 Transcriptome sequencing data quality statistics

Table 3 Statistics of high-expressed genes in wt and pmr1 (FPKM>3 000 means extremely high expression)

Table 4 Differentially expressed genes up-regulated in wt and pmr1Δ (log2FoldChange≥ 1 )

Table 5 Differentially expressed genes down-regulated in wt and pmr1Δ ( log2FoldChange≤ -1 )

Fig.5 Volcano plot of differentially expressed genes

Fig.6 GO enrichment of all differentially expressed genes

Fig.7 KEGG enrichment of all differentially expressed genes

Fig.8 Metabolic pathway enrichment of differentially expressed genes a) Glycolysis/Gluconeogenesis pathway (b) Fatty acid biosynthesis pathway (c) Mismatch repair pathway. Blue indicates down-regulated genes, red indicates up-regulated genes

Fig.9 qRT-PCR verification of key differentially expressed genes


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