Nucleolin from the multiple nucleoli of amphibian oocyte nuclei.
When fixed preparations of newt germinal vesicle (GV) contents are treated with RNase and are then probed with radiolabeled single-stranded DNA in 0.1-2.0 X SSC, the extrachromosomal nucleoli bind the probe non-specifically. DNA/protein blot analysis of proteins from newt GVs shows that gv95, an acidic protein (pI = 5.0) of Mr = 95,000, is the most prominent non-specific DNA-binding protein. Immunocytochemical analysis with affinity purified antibody directed against gv95 shows that it is located in the multiple nucleoli. We used an antibody directed against rat nucleolin to show that newt gv95 and two similar Xenopus GV proteins are the amphibian versions of nucleolin, a nucleolar ribonucleoprotein originally identified in mammalian cells. We show that mAb 3A10, directed against newt histones H1 and H5, labels gv95 on protein immunoblots and the multiple nucleoli in cytological preparations. These results suggest that histone H1 and nucleolin share a cross-reacting epitope.
DiMario
P J
PJ
Department of Embryology, Carnegie Institution of Washington, Baltimore, MD 21210.
Gall
J G
JG
D000663:Amphibians / Q000378:metabolism*; D000818:Animals; D002466:Cell Nucleolus / Q000032:analysis*; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006657:Histones / Q000276:immunology; D007150:Immunohistochemistry; D009673:Notophthalmus viridescens / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D009865:Oocytes / Q000032:analysis*; D010750:Phosphoproteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D016601:RNA-Binding Proteins; D014982:Xenopus laevis / Q000378:metabolism; D000097796:Nucleolin
D004247:DNA; D004268:DNA-Binding Proteins; D006657:Histones; D009687:Nuclear Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin