IMMUNOLOCALIZATION OF KU-PROTEINS (P80 P70) - LOCALIZATION OF P70 TO NUCLEOLI AND PERIPHERY OF BOTH INTERPHASE NUCLEI AND METAPHASE CHROMOSOMES
IMMUNOLOCALIZATION OF KU-PROTEINS (P80 P70) - LOCALIZATION OF P70 TO NUCLEOLI AND PERIPHERY OF BOTH INTERPHASE NUCLEI AND METAPHASE CHROMOSOMES
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DOI：
10.1016/0014-4827(92)90293-h
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发表时间：
1992-08-01
影响因子：
3.7
通讯作者：
YAGURA, T
中科院分区：
文献类型：
--
作者：
HIGASHIURA, M;SHIMIZU, Y;YAGURA, T
用特异性单克隆抗体（mAbH6）识别p70，一种Ku蛋白，研究了系统性红斑狼疮患者的自身抗体识别的Ku蛋白在细胞核和中期染色体上的分布。用常规荧光显微镜或共聚焦激光扫描显微镜观察发现，mAbH6染色的p70抗原定位于人间期细胞的核周边和核仁。用mAbH6对核仁的特异性染色已经使用从大鼠肝脏分离的核仁证实，其中染色被视为围绕核仁DNA的细颗粒。在有丝分裂过程中，p70抗原从与核膜区域的结合转移到凝聚染色体的外周上，而染色体内部没有明显的染色。使用mAbH6通过免疫亲和柱层析将p70抗原与DNA片段共纯化。用低浓度的DNA酶I消化后，核周边和核仁的mAbH6染色均消失。这些结果表明，p70抗原通过DNA与这些核结构连接。
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.