AI Article Synopsis
- The Cajal body (CB) is a crucial structure in the cell nucleus involved in important processes like RNA splicing and transcription complex assembly.
- Researchers developed a new method to purify CBs from HeLa cell nuclei, achieving a 750-fold increase in the CB marker protein p80-coilin.
- Analyses confirmed that the isolated CBs maintained similar structure and composition to those found naturally in cells, allowing for detailed studies on their molecular characteristics.
Article Abstract
The Cajal body (CB) is a conserved, dynamic nuclear structure that is implicated in various cellular processes, such as the maturation of splicing small nuclear ribonucleoproteins and the assembly of transcription complexes. Here, we report the first procedure for the large-scale purification of CBs from HeLa cell nuclei, resulting in an approximately 750-fold enrichment of the CB marker protein p80-coilin. Immunofluorescence, immunoblotting, and mass spectrometric analyses showed that the composition of the isolated CBs was similar to that of CBs in situ. The morphology and structure of the isolated CBs, as judged by transmission and scanning electron microscopy analysis, are also similar to those of CBs in situ. This protocol demonstrates the feasibility of isolating intact distinct classes of subnuclear bodies from cultured cells in sufficient yield and purity to allow detailed characterization of their molecular composition, structure, and properties.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC117327 | PMC |
| http://dx.doi.org/10.1091/mbc.02-03-0034 | DOI Listing |
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