Immunolocalization of Ku-proteins (p80/p70): localization of p70 to nucleoli and periphery of both interphase nuclei and metaphase chromosomes.
Distribution on both nuclei and metaphase chromosomes of Ku-proteins, recognized by autoantibodies from a patient with systemic lupus erythematosus, has been studied using a specific monoclonal antibody (mAbH6) that recognizes p70, one Ku-protein. Observation with either a conventional fluorescent microscope or a confocal laser scanning microscope revealed mAbH6-stained p70 antigen localized on both nuclear periphery and nucleoli of human interphase cells. The specific staining of nucleoli with mAbH6 has been confirmed using isolated nucleoli from rat liver in which the staining was seen as fine granules surrounding nucleolar DNA. During mitosis p70 antigen moved away from association with the nuclear envelope region to localization on the periphery of condensed chromosomes with no apparent staining of chromosome interior. The p70 antigen was copurified with DNA fragments by immunoaffinity column chromatography using mAbH6. The mAbH6 staining of both nuclear periphery and nucleoli was lost upon digestion with DNase I at low concentrations. These results suggest that p70 antigen is connected with these nuclear structures through DNA.
Higashiura
M
M
Department of Biology, Faculty of Science, Kwansei Gakuin University, Hyogo, Japan.
Shimizu
Y
Y
Tanimoto
M
M
Morita
T
T
Yagura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D002460:Cell Line; D002466:Cell Nucleolus / Q000737:chemistry*; D002875:Chromosomes / Q000737:chemistry*; D004265:DNA Helicases; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000187:drug effects / Q000276:immunology; D003850:Deoxyribonuclease I / Q000494:pharmacology; D006801:Humans; D007399:Interphase; D000072200:Ku Autoantigen; D008180:Lupus Erythematosus, Systemic / Q000378:metabolism; D008677:Metaphase; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000737:chemistry*; D009687:Nuclear Proteins / Q000032:analysis* / Q000276:immunology; D051381:Rats
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D003850:Deoxyribonuclease I; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; C000606511:Xrcc6 protein, rat; D000072200:Ku Autoantigen