Characterization of the 2H12 antigen as a nonshuttling human isoelectric variant of the nucleolar protein B23.
It has become obvious that a better understanding of the nucleolar compartment should encompass the elucidation of structural and functional relationships between its molecular constituents. Using a mouse monoclonal antibody referred to as 2H12, we have identified a human epitope that appears to be implicated in the regulatory events governing the elaboration and stabilization of the nucleolar architecture. By immunofluorescence and immunoblotting, the 2H12 monoclonal was shown to be directed against a nucleolar protein with a relative mobility of 38-40 kDa and an isoelectric point of 5.1 that is present in human cells, regardless of their proliferation state. No reactivity was detected in cells from other species, implying that the targeted epitope could be unique to humans. Investigation of the fate of the epitope throughout the cell cycle led to evidence that its immunoreactivity was phosphodependent and suggested that the disassembly and reassembly of the nucleolar apparatus during cell division is accompanied by dephosphorylation/phosphorylation modifications at this site. In a series of double immunofluorescence experiments and two-dimensional immunoblotting analyses, it was demonstrated that the 2H12 antigen corresponds to an isoelectric variant of the human nucleolar protein B23 that is most prominent during interphase. Tightly associated with the nuclear matrix, this human B23 isoelectric variant did not shuttle between the nucleus and the cytoplasm but remained sequestered within the human nucleolus during mobility assays in human-murine heterokaryons.
Paulin-Levasseur
M
M
Department of Biology, University of Ottawa, Ontario, Canada.
Julien
M
M
Horner
M
M
Chen
G
G
D000818:Animals; D000941:Antigens / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D002459:Cell Fusion; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D007526:Isoelectric Point; D051379:Mice; D009687:Nuclear Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D000090243:Nucleophosmin; D051381:Rats; D013045:Species Specificity
D000941:Antigens; D000939:Epitopes; C000715570:Npm1 protein, mouse; D009687:Nuclear Proteins; D000090243:Nucleophosmin