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

Abstract

Objective: The pmrl 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 pmrl 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 pmrl deletion. Methods: The effects of pmrl deletion on cell mitosis and sexual reproduction were detected by cell growth rate measurement, spore production observation and statistics, green fuorescent protein-labeled monitoring and living cell imaging; The wild-type and pmrl A strain were sequenced by RNA-Seq and analyzed by bioinformatics, and verified by qRT-PCR. Results: The pmrl 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 mfml, mfm2 and matl -Mc, up-regulation of cdcl and exol in the mismatch repair pathway, and down-regulation of pgil, pfkl and dldl in the glycolysis/gluconeogenesis pathway are the main factor of the increased spore length in the pmrl A; Meanwhile, the down-regulation of fdhl and pgkl in the glycolysis/gluconeogenesis pathway and the down-regulation of fasl, fas2, cut6 and Icfl in the fatty acid anabolic pathway also led to the decreased cell length in mitosis of pmrl A; The up-regulation of hsp9 is the key gene that affected the formation time of actin ring in pmrl A. qRT-PCR experiments confirmed that the expression trends of key genes after pmrl deletion were consistent with the RNA-Seq results. Conclusion: After pmrl 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.