Institute of Genome Sciences, School of Life Sciences, National Yang-Ming University, Taipei, Taiwan.
PLoS One. 2012;7(3):e32820. doi: 10.1371/journal.pone.0032820. Epub 2012 Mar 5.
In this study, we employed a surface-specific antibody against the large ribosome subunit to investigate the distribution of ribosomes in cells during the cell cycle. The antibody, anti-L7n, was raised against an expansion segment (ES) peptide from the large subunit ribosomal protein L7, and its ribosome-surface specificity was evident from the positive immuno-reactivity of ribosome particles and the detection of 60 S immune-complex formation by an immuno-electron microscopy. Using immunofluorescent staining, we have microscopically revealed that ribosomes are dispersed in the cytoplasm of cells throughout all phases of the cell cycle, except at the G2 phase where ribosomes show a tendency to gather toward the nuclear envelope. The finding in G2 cells was confirmed by electron microscopy using a morphometric assay and paired t test. Furthermore, further observations have shown that ribosomes are not distributed immune-fluorescently with nuclear envelope markers including the nuclear pore complex, the integral membrane protein gp210, the inner membrane protein lamin B2, and the endoplasm reticulum membrane during cell division we propose that the mechanism associated with ribosome segregation into daughter cells could be independent of the processes of disassembly and reassembly of the nuclear envelope.
在这项研究中,我们使用了一种针对大亚基的表面特异性抗体来研究细胞周期中核糖体在细胞中的分布。该抗体抗-L7n 是针对核糖体大亚基蛋白 L7 的一个扩展片段 (ES) 肽产生的,其核糖体表面特异性从核糖体颗粒的阳性免疫反应和免疫电子显微镜检测到的 60S 免疫复合物形成中明显看出。使用免疫荧光染色,我们通过显微镜揭示了核糖体在细胞周期的所有阶段都分散在细胞质中,除了在 G2 期,核糖体显示出向核膜聚集的趋势。这一发现在 G2 细胞中通过电子显微镜使用形态计量测定和配对 t 检验得到了证实。此外,进一步的观察表明,核糖体在细胞分裂过程中与核膜标记物(包括核孔复合体、整膜蛋白 gp210、内膜蛋白 lamin B2 和内质网膜)没有免疫荧光分布,我们提出与核糖体分离到子细胞相关的机制可能独立于核膜的拆卸和重新组装过程。