Molecular cloning of metaphase chromosome protein 1 (MCP1), a novel human autoantigen that associates with condensed chromosomes during mitosis
Molecular cloning of metaphase chromosome protein 1 (MCP1), a novel human autoantigen that associates with condensed chromosomes during mitosis
复制标题
DOI：
10.1023/a:1009230811398
复制
发表时间：
1998-04-01
影响因子：
2.6
通讯作者：
Sunkel, CE
中科院分区：
文献类型：
--
作者：
Bronze-da-Rocha, E;Catita, JA;Sunkel, CE
系统性红斑狼疮自身抗体被用来识别和表征新的人类染色体相关蛋白。先前在人类和小鼠组织培养细胞中的免疫定位研究表明，这些单克隆抗体中的一些识别在有丝分裂期间与浓缩的染色体相关联的核抗原。选择一种抗体用于筛选人HeLa S3 cDNA表达文库，并分离编码31-33 kDa抗原的cDNA。免疫学、生物化学和细胞分级数据表明，31- 33-kDa抗原对应于由原始单克隆抗体识别的染色体相关蛋白。序列分析表明，我们分离到一个新的人类基因。对人组织培养细胞的免疫定位显示，在间期，抗原分散在细胞核中，在有丝分裂期间，它只与浓缩的染色体相关联。在小鼠成纤维细胞中也观察到类似的定位模式，表明该抗原在不同物种中是保守的。最后，我们表明，即使在高盐提取后，部分抗原仍与支架/基质组分结合。
Systemic lupus erythematosus autoantibodies were used to identify and to characterize new human chromosome-associated proteins. Previous immunolocalization studies in human and murine tissue culture cells showed that some of these monoclonal antibodies recognize nuclear antigens that associate with condensed chromosomes during mitosis. One antibody was selected for screening a human HeLa S3 cDNA expression library, and cDNAs that code for an antigen of 31-33 kDa were isolated. Immunological, biochemical and cell fractionation data indicate that the 31- to 33-kDa antigen corresponds to the chromosome-associated protein recognized by the original monoclonal antibody. Sequence analysis shows that we isolated a novel human gene. Immunolocalization to human tissue culture cells shows that during interphase the antigen is dispersed in the nucleus and that during mitosis it associates exclusively with condensed chromosomes. A similar pattern of localization was also observed in mouse fibroblasts, suggesting that the antigen is conserved among different species. Finally, we show that part of the antigen remains bound to the scaffold/matrix component, even after high salt extraction.