Abstract:
The rpl1001 gene encodes 60S ribosomal protein L10, which is involved in intracellular protein synthesis and cell growth. However, it is not yet known whether it is involved in the regulation of cell mitosis dynamics. This study focuses on the growth, spore production, cell morphology, the dynamics of microtubules, chromosomes, actin, myosin, and mitochondria of fission yeast (Schizosaccharomyces pombe) to investigate the impact of rpl1001 deletion on cell mitosis. RNA-Seq and bioinformatics analyses were also used to reveal key genes, such as hsp16, mfm1 and isp3, and proteasome pathways. The results showed that rpl1001 deletion resulted in slow cell growth, abnormal spore production, altered cell morphology, and abnormal microtubule number and length during interphase. The cell dynamics of the rpl1001Δ strain showed that the formation of a monopolar spindle leads to abnormal chromosome segregation with increased rate of spindle elongation in anaphase of mitosis, decreased total time of division, prolonged formation time of actin and myosin loops, and increased expression of mitochondrial proteins. Analysis of the RNA-Seq sequencing results showed that the proteasome pathway, up-regulation of isp3, and down-regulation of mfm1 and mfm2 in the rpl1001Δ strain were the main factors underpinning the increased number of spore production. Also, in the rpl1001Δ strain, down-regulation of dis1 caused the abnormal microtubule and chromosome dynamics, and down-regulation of hsp16 and pgk1 were the key genes affecting the delay of actin ring and myosin ring formation. This study reveals the effect and molecular mechanism of rpl1001 gene deletion on cell division, which provides the scientific basis for further clarifying the function of the Rpl1001 protein in cell division.
摘要:
rpl1001基因编码60S核糖体蛋白L10,其参与细胞内蛋白质合成和细胞生长。然而,目前尚不清楚它是否参与细胞有丝分裂动力学的调节。这项研究的重点是增长,孢子生产,细胞形态学,微管的动力学,染色体,肌动蛋白,肌球蛋白,和裂殖酵母(裂殖酵母)的线粒体,以研究rpl1001缺失对细胞有丝分裂的影响。RNA-Seq和生物信息学分析也用于揭示关键基因,如hsp16,mfm1和isp3,以及蛋白酶体途径。结果表明,rpl1001缺失导致细胞生长缓慢,孢子产生异常,改变细胞形态,间期微管数量和长度异常。rpl1001Δ菌株的细胞动力学表明,单极纺锤体的形成导致染色体异常分离,有丝分裂后期纺锤体伸长率增加,减少了总的分裂时间,肌动蛋白和肌球蛋白环的形成时间延长,线粒体蛋白的表达增加。RNA-Seq测序结果表明,蛋白酶体通路,rpl1001Δ菌株中isp3的上调和mfm1和mfm2的下调是支持孢子产量增加的主要因素。此外,在rpl1001Δ应变中,dis1的下调导致微管和染色体动力学异常,hsp16和pgk1的下调是影响肌动蛋白环和肌球蛋白环形成延迟的关键基因。本研究揭示了rpl1001基因缺失对细胞分裂的影响及其分子机制。为进一步阐明Rpl1001蛋白在细胞分裂中的功能提供了科学依据。