Effect of uge1 gene deletion on the growth and reproduction of Schizosaccharomyces pombe and its molecular mechanism

doi:10.3969/j.issn.1000-1565.2025.06.008

Abstract

Abstract: In order to clarify the reproductive status, dynamic changes in mitosis interphase and the abnormal key expression genes and key metabolic pathways after uge1 gene deletion of Schizosaccharomyces pombe, live cell imaging was used to analyze sporulation and dynamic changes in mitosis interphase after uge1 gene deletion(Δuge1)combining with RNA sequencing(RNA-Seq)technology and bioinformatics analysis for wild-type and Δuge1 strain, and the expression of key genes was- DOI:10.3969/j.issn.1000-1565.2025.06.008uge1基因缺失对粟酒裂殖酵母生长生殖的影响及其分子机制青昕¹,张丁¹,余雯¹,刘可¹,丁祥²,侯怡铃¹(1.西华师范大学生命科学学院,西南野生动植物资源保护教育部重点实验室,四川南充 637009;2.西华师范大学环境科学与工程学院,四川南充 637009)摘要:为明确粟酒裂殖酵母uge1基因缺失后生殖情况、有丝分裂间期动力学变化及异常的关键表达基因和关键代谢通路,采用活细胞成像分析uge1基因缺失(Δuge1)后产孢情况及有丝分裂间期动力学变化,结合RNA测序(RNA-Seq)技术对野生型和Δuge1菌株测序并进行生物信息学分析,同时采用荧光定量PCR对关键基因的表达水平进行验证. 活细胞成像结果表明,uge1基因缺失导致酵母产4个正常孢子数的细胞减少,孢子直径增大,6%的Δuge1菌株出现细胞核解聚,微管束在细胞壁上的停留时间减少了(0.526±0.08)min. RNA测序结果显示,Δuge1菌株中mat2-Pc、mat2-Pi基因的上调表达与mfm1、mfm2基因的下调表达导致了粟酒裂殖酵母的异常生殖过程,aat1基因的表达量增加使纺锤体延伸时间变长,acp2和cdc8基因的过量表达则导致了肌动蛋白环收缩减慢. KEGG富集分析表明,脂肪酸合成代谢中fas1、fas2和cut6基因的表达下调干扰了脂肪酸合成代谢,促使核膜合成障碍,导致核解聚的发生. 研究结果揭示了粟酒裂殖酵母uge1基因缺失导致细胞生长生殖异常的关键表达基因和关键代谢通路,为深入研究uge1基因对细胞动力学影响的分子机制提供了一定的科学依据.关键词:uge1基因缺失;粟酒裂殖酵母;RNA测序;有性生殖;有丝分裂中图分类号:Q932 文献标志码:A 文章编号:1000-1565(2025)06-0623-14Effect of uge1 gene deletion on the growth and reproduction of Schizosaccharomyces pombe and its molecular mechanismQING Xin¹, ZHANG Ding¹, YU Wen¹, LIU Ke¹, DING Xiang², HOU Yiling¹(1. Key Laboratory of Southwest China Wildlife Resources Conservation, the 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)Abstract: In order to clarify the reproductive status, dynamic changes in mitosis interphase and the abnormal key expression genes and key metabolic pathways after uge1 gene deletion of Schizosaccharomyces pombe, live cell imaging was used to analyze sporulation and dynamic changes in mitosis interphase after uge1 gene deletion(Δuge1)combining with RNA sequencing(RNA-Seq)technology and bioinformatics analysis for wild-type and Δuge1 strain, and the expression of key genes was- 收稿日期:2024-10-30;修回日期:2025-03-03 基金项目:四川省科技厅应用基础面上项目(2022NSFSC0107);四川省科技厅农业科技成果转化资金项目(2022NZZJ0003);四川省转移支付科技成果转移转化示范项目(22ZYZFSF0009;23ZHSF0082) 第一作者:青昕(1998—),女,西华师范大学在读硕士研究生,主要从事生物化学与分子生物学方向研究.E-mail:1026362669@qq.com 通信作者:侯怡铃(1983—),女,西华师范大学教授,主要从事生物化学与分子生物学方向研究.E-mail:yilingh128@163.com 第6期青昕等:uge1基因缺失对粟酒裂殖酵母生长生殖的影响及其分子机制河北大学学报(自然科学版) 第45卷also validated using fluorescence quantitative PCR. The results of live cell imaging showed that the deletion of the uge1 gene resulted in a decrease in the number of cells producing four normal spores in yeast, an increase in spore diameter. 6% Δuge1 strain exhibited nuclear depolymerization and the residence time of the microtubule bundle on the cell wall decreased by(0.526±0.08)min. RNA sequencing results showed that the up-regulated expression of mat2-Pc and mat2-Pi genes and the down-regulation of mfm1 and mfm2 genes in the Δuge1 strain led to the abnormal reproductive process of S.pombe. The increased expression of aat1 gene interfered with the extension of spindle, and the overexpression of acp2 and cdc8 genes led to the slow contraction of actin loop in Δuge1 strain. KEGG enrichment analysis showed that the down-regulation of fas1, fas2 and cut6 genes in fatty acid synthesis pathway interfered with the fatty acid synthesis, which promoted the disorder of nuclear membrane synthesis, and led to the occurrence of nuclear depolymerization. These results revealed the key genes and key metabolic pathways that the deletion of uge1 gene in S.pombe led to abnormal cell growth and reproduction, and provided a scientific basis for further research on the molecular mechanism of uge1 genes impact on cell dynamics.

Key words: uge1 gene deletion, Schizosaccharomyces pombe, RNA-Seq, sexual reproduction, mitosis

CLC Number:

Q932

QING Xin, ZHANG Ding, YU Wen, LIU Ke, DING Xiang, HOU Yiling. Effect of uge1 gene deletion on the growth and reproduction of Schizosaccharomyces pombe and its molecular mechanism[J]. Journal of Hebei University(Natural Science Edition), 2025, 45(6): 623-636.

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