Abstract
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.
Highlights
The biogenesis of a ribosome in the eukaryotic cell can be detected at the start cell cycle checkpoint [1], and it involves many aspects of the cellular machinery [2]
In addition to the above, the ribosome-surface specificity of the anti-L7n antibody was further ratified by an immuno-electron microscopy (IEM) [16] where the formation of 60 S immunecomplexes (60 S dimers) (Figs. 1D and 1E) was confirmed after incubation of the 60 S subunit with anti-L7n antibody
This revealed the relationship between ribosome distribution and the nuclear envelope during each phase of the cell cycle
Summary
The biogenesis of a ribosome in the eukaryotic cell can be detected at the start cell cycle checkpoint [1], and it involves many aspects of the cellular machinery [2]. To understand how a cell can command ribosome movement in cytoplasm to allow translation is of significant interest. How a cell distributes its ribosome particles during the cell cycle is important. The latter issue would have a great impact on the survival of the daughter cells, which need an adequate number of ribosomes to ensure the synthesis of important proteins for future physiological events [8]. These issues have gone unstudied because, as suggested earlier [9], there is a lack of a good method for pinpointing and counting the ribosome particles in the cell. Using immunofluorescent staining by a specific ribosome-surface antibody would be an ideal tool for localizing ribosome particles during cellular events, but such an antibody is quite difficult to produce
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.