Characteristic immunolocalization of Ku protein as nuclear matrix.
Two hybridoma clones, NMB1 and NML90, were established using nuclear matrix proteins from normal human thymi or malignant lymphoma as immunogens. They reacted with human Ku70 and Ku80, respectively, by immunoblotting. When HeLa cell nuclear proteins were fractionated and applied to immunoblotting, both Ku70 and Ku80 were detected in the nuclear matrix as well as the soluble nuclear protein fractions. By confocal scanning microscopy, the immunoreactivity of Ku70 and Ku80 was localized to distinct nucleoplasmic fibrillar network and fine granules in the interphase cell nuclei. When HeLa cells were fractionated in situ using DNase I and buffers containing 0.25 M (NH4)2SO4 and 2 M NaCl, the nucleoplasmic reticular structure was largely preserved, but granules disappeared. The nucleoplasmic distribution of Ku in the tissue and in cultured cells was distinct from each other. In the adult tissue, it consisted mostly of either distinct curvilinear lines along the nuclear periphery or of tangled, beaded lines throughout the nuclei. When xenotransplants of HeLa cell in Scid mice were examined, the "tissue type" immunolocalization pattern was reproduced consistently. In most fetal tissues, "tissue type" and "cell type" patterns were admixed. Monoclonal antibodies described here are useful tools for studying the structure and function of the nuclear matrix.
Yu
E
E
Department of Pathology, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.
Song
K
K
Moon
H
H
Maul
G G
GG
Lee
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D002478:Cells, Cultured; D004265:DNA Helicases; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas; D015151:Immunoblotting; D000072200:Ku Autoantigen; D008099:Liver / Q000196:embryology / Q000648:ultrastructure; D008113:Liver Neoplasms / Q000648:ultrastructure; D008297:Male; D051379:Mice; D016513:Mice, SCID; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015530:Nuclear Matrix / Q000737:chemistry* / Q000276:immunology; D009687:Nuclear Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011233:Precipitin Tests; D013093:Spermatogonia / Q000648:ultrastructure; D013094:Spermatozoa / Q000648:ultrastructure; D013737:Testis / Q000648:ultrastructure; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; C000606510:Xrcc6 protein, mouse; D000072200:Ku Autoantigen