The dynamic organization of the perinucleolar compartment in the cell nucleus

Huang, S., Deerinck, T. J., Ellisman, M. H. , Spector, D. L. (1997) The dynamic organization of the perinucleolar compartment in the cell nucleus. Journal of Cell Biology, 137 (5). pp. 965-74. ISSN 0021-9525

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URL: http://www.ncbi.nlm.nih.gov/pubmed/9166399
DOI: 10.1083/jcb.137.5.965

Abstract

The perinucleolar compartment (PNC) is a unique nuclear structure preferentially localized at the periphery of the nucleolus. Several small RNAs transcribed by RNA polymerase III (e.g., the Y RNAs, MRP RNA, and RNase P H1 RNA) and the polypyrimidine tract binding protein (PTB; hnRNP I) have thus far been identified in the PNC (Ghetti, A., S. PinolRoma, W.M. Michael, C. Morandi, and G. Dreyfuss. 1992. Nucleic Acids Res. 20:3671-3678; Matera, A.G., M.R. Frey, K. Margelot, and S.L. Wolin. 1995. J. Cell Biol. 129:1181-1193; Lee, B., A.G. Matera, D.C. Ward, and J. Craft. 1996. Proc. Natl. Acad. Sci. USA. 93: 11471-11476). In this report, we have further characterized this structure in both fixed and living cells. Detection of the PNC in a large number of human cancer and normal cells showed that PNCs are much more prevalent in cancer cells. Analysis through the cell cycle using immunolabeling with a monoclonal antibody, SH54, specifically recognizing PTB, demonstrated that the PNC dissociates at the beginning of mitosis and reforms at late telophase in the daughter nuclei. To visualize the PNC in living cells, a fusion protein between PTB and green fluorescent protein (GFP) was generated. Time lapse studies revealed that the size and shape of the PNC is dynamic over time. In addition, electron microscopic examination in optimally fixed cells revealed that the PNC is composed of multiple strands, each measuring approximately 80-180 nm diam. Some of the strands are in direct contact with the surface of the nucleolus. Furthermore, analysis of the sequence requirement for targeting PTB to the PNC using a series of deletion mutants of the GFP-PTB fusion protein showed that at least three RRMs at either the COOH or NH2 terminus are required for the fusion protein to be targeted to the PNC. This finding suggests that RNA binding may be necessary for PTB to be localized in the PNC.

Item Type: Paper
Uncontrolled Keywords: Adenocarcinoma Amino Acid Sequence Biological Transport physiology Breast Neoplasms Carcinoma Ductal Breast Cell Compartmentation physiology Cell Cycle physiology Cell Line Transformed Cell Nucleolus physiology ultrastructure Colonic Neoplasms Comparative Study DNA-Binding Proteins chemistry genetics pharmacokinetics Female Fibroblasts cytology physiology ultrastructure Green Fluorescent Proteins Hela Cells Humans Luminescent Proteins pharmacokinetics Lung cytology Microscopy Electron Mutagenesis physiology Phenotype Polypyrimidine Tract Binding Protein RNA-Binding Proteins chemistry genetics pharmacokinetics Recombinant Fusion Proteins pharmacokinetics Research Support US Govt PHS Skin cytology
Subjects: organs, tissues, organelles, cell types and functions > organelles, types and functions > nucleus
organs, tissues, organelles, cell types and functions > tissues types and functions
CSHL Authors:
Communities: CSHL labs > Spector lab
Depositing User: Brian Soldo
Date Deposited: 04 Apr 2012 19:05
Last Modified: 29 Jan 2015 17:42
PMCID: PMC2136227
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25663

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