sgf73基因敲除对粟酒裂殖酵母细胞有丝分裂的影响  

Effect of sgf73 Gene Knockout during Mitosis of Schizosaccharomyces pombe Cells

作　　者：袁荣美 丁祥 谭秀梅 鲁艳 周丽倩 刘欣岚 侯怡铃[1] YUAN Rong-mei;DING Xiang;TAN Xiu-mei;LU Yan;ZHOU Li-qian;LIU Xin-lan;HOU Yi-ling(Key Lab.of SW China Wildlife Res.Conserv.,Minist.of Educ.,Coll.of Life Sci.,W China Normal Uni.,Nanchong 637009;Sichuan Ctr.for Dis.Ctrl.&Prevent.,Chengdu 610044;Coll.of Environ.Sci.&Engin.,W China Normal Uni.,Nanchong 637009;Managem′t Comm.of Nanxi Econ.Devel.Zone,Yibin,Nanxi 644100)

机构地区：[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]四川省疾病预防控制中心,四川成都610044 [3]西华师范大学环境科学与工程学院,四川南充637009 [4]四川宜宾南溪经济开发区管理委员会,四川南溪644100

出　　处：《微生物学杂志》2022年第6期10-24,共15页Journal of Microbiology

基　　金：四川省科技厅应用基础重点项目(2018JY0087);四川省科技厅重点研发项目(2018NZ0055)。

摘　　要：sgf73基因编码的Sgf73蛋白对SAGA复合物的功能具有重要作用。为探究sgf73基因缺失后sgf73Δ菌株有丝分裂中纺锤体、染色体、肌动蛋白的动力学变化,以粟酒裂殖酵母(Schizosaccharomyces pombe)为材料,对sgf73Δ菌株进行生长曲线和减数分裂产孢实验;采用荧光蛋白标记和活细胞成像的方法,对sgf73Δ菌株有丝分裂进行观察。生长曲线结果分析发现,sgf73基因敲除极显著影响粟酒裂殖酵母的生长,并导致子囊孢子数目显著减少。活细胞成像结果分析发现,在有丝分裂间期,sgf73Δ菌株微管数量显著增加、微管长度趋于增长但无显著差异;sgf73Δ菌株在分裂期纺锤体出现组装缺陷,单极纺锤体极显著增加,前期和中期纺锤体伸长速率显著下降、后期纺锤体伸长速率极显著下降、分裂中期和后期时间分别显著和极显著延长,有丝分裂总时间极显著延长,细胞长度也增长。肌动蛋白分裂环在有丝分裂中维持及收缩时间出现极显著和显著延长,总速率显著降低。同时,染色体分离出现缺陷。以上结果表明,sgf73基因缺失会导致菌株微管、染色体、肌动蛋白缺陷。Sgf73 protein encoded by sgf73 gene plays an important role in the function of SAGA complex.In order to explore the kinetic changes of spindle,chromosome and actin during mitosis of sgf73Δcells,the fission yeast(Schizosaccharomyces pombe)was used as material to study the growth curve and meiotic sporogenesis of sgf73Δstrain and fluorescent protein labeling and living cell imaging were used to detect the mitosis of sgf73Δstrains.The results of growth curve analysis showed that sgf73 gene knockout very significantly affected the growth of sgf73Δstrains and significantly reduced the number of ascospores.The analysis of living cell imaging results showed that the number of microtubules of sgf73Δstrains was increased significantly and the length of microtubules tended to increase,but no significant difference;during the mitotic stage,the spindle assembly defect occurred,the monopolar spindle increased very significantly,the spindle elongation rate decreased significantly in prophase and metaphase,decreased significantly in anaphase.The time of metaphase and anaphase significantly and extremely significantly prolonged,respectively,the total mitotic time was prolonged significantly,and cell length also increased.The maintenance and contraction time of actin ring in mitosis increased very significantly and significantly,respectively,and the total rate decreased significantly.At the same time,chromosome segregation also defective.These results indicated that the deletion of sgf73 gene can led to the defects of microtubule,chromosome and actin.

关 键 词：粟酒裂殖酵母 sgf73 基因敲除 有丝分裂 细胞动力学 

分 类 号：Q933[生物学—微生物学]