Molecular cloning of metaphase chromosome protein 1 (MCP1), a novel human autoantigen that associates with condensed chromosomes during mitosis.
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.
Bronze-da-Rocha
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Laboratório de Genética Molecular, Instituto de Biologia Molecular e Celular da Universidade do Porto, Portugal.
Catita
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Sunkel
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CE
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001324:Autoantigens / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002457:Cell Extracts; D002460:Cell Line; D002877:Chromosomes, Human / Q000737:chemistry*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018628:Gene Dosage; D006367:HeLa Cells; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008677:Metaphase; D051379:Mice; D008938:Mitosis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D002457:Cell Extracts; D018076:DNA, Complementary; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C487774:metaphase chromosome protein 1, human