Subcellular Distribution of Distinct Nucleolin Subfractions Recognized by Two Monoclonal Antibodies

Monoclonal antibodies binding to different domains of nucleolin have been used to localize nucleolin in tissue culture cells ofXenopus laevis.The monoclonal antibody b6-6E7 binds to an epitope in the N-terminal domain, which contains arrays of phosphorylation consensus sites. This monoclonal antibod...

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Bibliographic Details
Published in:Experimental cell research 1998-03, Vol.239 (2), p.226-234
Main Authors: Schwab, Markus S., Goβweiler, Ulrike, Dreyer, Christine
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - immunology
Antibody Specificity
Cell Compartmentation
Cell Cycle
Cell Nucleolus - chemistry
Cells, Cultured
Epithelial Cells - chemistry
Epithelial Cells - ultrastructure
Epitopes - immunology
Fluorescent Antibody Technique, Indirect
Kidney
Larva - cytology
Meiosis
Metaphase
Mitosis
Nucleolin
Oocytes - chemistry
Oocytes - ultrastructure
Phosphoproteins - analysis
Phosphoproteins - chemistry
Phosphoproteins - immunology
Ribonucleases - pharmacology
RNA, Ribosomal - metabolism
RNA-Binding Proteins - analysis
RNA-Binding Proteins - chemistry
RNA-Binding Proteins - immunology
Subcellular Fractions
Xenopus laevis
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Summary:Monoclonal antibodies binding to different domains of nucleolin have been used to localize nucleolin in tissue culture cells ofXenopus laevis.The monoclonal antibody b6-6E7 binds to an epitope in the N-terminal domain, which contains arrays of phosphorylation consensus sites. This monoclonal antibody binds to nucleolin of oocytes and of eggs with high affinity. In contrast, the monoclonal antibody Nu-1H6 binds poorly to the modified forms of nucleolin arising during meiosis and mitosis. In interphase cells, monoclonal antibody b6-6E7 preferentially stains the periphery of the nucleoli, where most of the rRNA accumulates. Staining by monoclonal antibody Nu-1H6 complements this pattern by staining mainly the center of the nucleoli. The epitope of monoclonal antibody Nu-1H6 is within the central domain of nucleolin, which contains the first two RNA binding domains. RNase treatment of cells results in loss of nucleolin from nucleoli. In mitotic cells, both monoclonal antibodies decorate the surface of condensing chromosomes in prophase. The periphery of the condensed chromosomes in metaphase and anaphase is preferentially stained by monoclonal antibody b6-6E7.
ISSN:0014-4827
1090-2422
DOI:10.1006/excr.1997.3878