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Proteomics analysis of the centromere complex from HeLa interphase cells: UV‐damaged DNA binding protein 1 (DDB‐1) is a component of the CEN‐complex, while BMI‐1 is transiently co‐localized with the centromeric region in interphase
Author(s) -
Obuse Chikashi,
Yang Hua,
Nozaki Naohito,
Goto Shouhei,
Okazaki Tuneko,
Yoda Kinya
Publication year - 2004
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2004.00705.x
Subject(s) - centromere , chromatin , biology , interphase , chromatin immunoprecipitation , microbiology and biotechnology , histone , histone h3 , chromosome , genetics , dna , gene , gene expression , promoter
CENP‐A, a centromere‐specific histone H3, is conserved throughout eukaryotes, and formation of CENP‐A chromatin defines the active centromere region. Here, we report the isolation of CENP‐A chromatin from HeLa interphase nuclei by chromatin immunoprecipitation using anti‐CENP‐A monoclonal antibody, and systematic identification of its components by mass spectrometric analyses. The isolated chromatin contained CENP‐B, CENP‐C, CENP‐H, CENP‐I/hMis 6 and hMis 12 as well as CENP‐A, suggesting that the isolated chromatin may represent the centromere complex (CEN‐complex). Mass spectrometric analyses of the CEN‐complex identified approximately 40 proteins, including the previously reported centromere proteins and the proteins of unknown function. In addition, we unexpectedly identified a series of proteins previously reported to be related to functions other than chromosome segregation, such as uvDDB‐1, XAP8, hSNF2H, FACTp180, FACTp80/SSRP1, polycomb group proteins (BMI‐1, RING1, RNF2, HPC3 and PHP2), KNL5 and racGAP. We found that uvDDB‐1 was actually localized to the centromeric region throughout cell cycle, while BMI‐1 was transiently co‐localized with the centromeres in interphase. These results give us new insights into the architecture, dynamics and function of centromeric chromatin in interphase nuclei, which might reflect regulation of cell proliferation and differentiation.