Article: Human CENP-H multimers colocalize with CENP-A and CENP-C at active centromere--kinetochore complexes.
2000 Volume 9, Issue 19, Page(s) 2919–2926
Abstract: ... CENP-A and CENP-C in both interphase and metaphase. CENP-H was present outside centromeric ... that the CENP-H binds to itself and MCAK, but not to CENP-A, CENP-B or CENP-C. CENP-H multimers were observed ... the isolation and characterization of human CENP-H. Confocal microscopic analyses of HeLa cells with anti-human ...
| Abstract | Centromere and kinetochore proteins have a pivotal role in centromere structure, kinetochore formation and sister chromatid separation. However, the molecular architecture and the precise dynamic function of the centromere-kinetochore complex during mitosis remain poorly understood. Here we report the isolation and characterization of human CENP-H. Confocal microscopic analyses of HeLa cells with anti-human CENP-H-specific antibody demonstrated that CENP-H colocalizes with inner kinetochore plate proteins CENP-A and CENP-C in both interphase and metaphase. CENP-H was present outside centromeric heterochromatin, where CENP-B is localized, and inside the kinetochore corona, where CENP-E is localized during prometaphase. Furthermore, CENP-H was detected at neocentromeres, but not at inactive centromeres in stable dicentric chromosomes. In vitro binding assays of human CENP-H with centromere-kinetochore proteins suggest that the CENP-H binds to itself and MCAK, but not to CENP-A, CENP-B or CENP-C. CENP-H multimers were observed in cells in which both FLAG-tagged CENP-H and hemagglutinin-tagged CENP-H were expressed. These results suggest that CENP-H multimers localize constitutively to the inner kinetochore plate and play an important fundamental role in organization and function of the active human centromere-kinetochore complex. |
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| MeSH term(s) | Amino Acid Sequence ; Autoantigens ; Centromere/metabolism ; Centromere Protein A ; Chromosomal Proteins, Non-Histone/chemistry ; Chromosomal Proteins, Non-Histone/genetics ; Chromosomal Proteins, Non-Histone/metabolism ; Cloning, Molecular ; Fluorescent Antibody Technique ; HeLa Cells ; Humans ; In Situ Hybridization, Fluorescence ; Kinetochores/metabolism ; Macromolecular Substances ; Microscopy, Fluorescence ; Mitosis ; Molecular Sequence Data ; Protein Binding ; Recombinant Fusion Proteins/metabolism ; Sequence Alignment |
| Chemical Substances | Autoantigens ; CENPA protein, human ; CENPH protein, human ; Centromere Protein A ; Chromosomal Proteins, Non-Histone ; Macromolecular Substances ; Recombinant Fusion Proteins ; centromere protein C |
| Language | English |
| Publishing date | 2000-11-22 |
| Publishing country | England |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. |
| ZDB-ID | 1108742-0 |
| ISSN | 1460-2083 ; 0964-6906 |
| ISSN (online) | 1460-2083 |
| ISSN | 0964-6906 |
| DOI | 10.1093/hmg/9.19.2919 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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