KINETOCHORE COMPONENTS RECOGNIZED BY HUMAN AUTOANTIBODIES ARE PRESENT ON MONONUCLEOSOMES
KINETOCHORE COMPONENTS RECOGNIZED BY HUMAN AUTOANTIBODIES ARE PRESENT ON MONONUCLEOSOMES
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DOI：
10.1128/mcb.5.1.173
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发表时间：
1985-01-01
影响因子：
5.3
通讯作者：
MARGOLIS, RL
中科院分区：
文献类型：
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作者：
PALMER, DK;MARGOLIS, RL
采用人钙质沉着症、雷诺现象、食道功能障碍、硬结症、毛细血管扩张症等硬皮病自身免疫抗体，建立了一种具有竞争性的酶联免疫吸附法检测溶解着丝点成分。在微球菌核酸酶消化的大鼠肝脏和鸡红细胞核的低盐和中高盐（375 mM盐）提取物中发现了间期持久成分或着丝粒前成分。免疫荧光显微镜显示，在这些条件下，细胞核中抗原活性的释放与着丝点前病灶的丢失有关。鸡红细胞核提取物的生化和竞争分析表明，着丝粒前组分与单个核小体大小的染色质紧密结合。基于竞争分析数据的结论得到了直接结合试验的支持，该试验证实CREST血清识别的抗原存在于染色质上。检测到的着丝酶特异性染色体抗原可替代标准的单核小体成分，如组蛋白H1。提出了从高等真核生物中分离着丝酶特异性DNA序列的方法。
A competitive enzyme-linked immunosorbent assay was developed for solubilized kinetochore components, using human CREST (calcinosis, Raynaud''s phenomenon, esophageal dysfunction, sclerodactyly, telangiectasia) scleroderma autoimmune antibodies specific for these kinetochore elements. Interphase persistent or pre-kinetochore components were found in low- and moderately high-salt (375 mM salt) extracts of micrococcal nuclease-digested rat liver and chicken erythrocyte nuclei. The release of antigen activity from nuclei under these conditions was correlated with loss of pre-kinetochore foci as determined by immunofluorescence microscopy. Combined biochemical and competition assay analysis of chicken erythrocyte nuclear extracts indicates that pre-kinetochore components are tightly bound to chromatin of mononucleosome size. The conclusions based on competition assay data are supported by a direct binding assay, which confirms that antigens recognized by CREST sera are present on chromatin. The kinetochore-specific chromosomal antigen(s) detected may substitute for standard mononucleosome components, such as histone H1. Approaches to the isolation of kinetochore-specific DNA sequences from higher eukaryotes are suggested.