CENP-B interacts with CENP-C domains containing Mif2 regions responsible for centromere localization.
Recently, human artificial chromosomes featuring functional centromeres have been generated efficiently from naked synthetic alphoid DNA containing CENP-B boxes as a de novo mechanism in a human cultured cell line, but not from the synthetic alphoid DNA only containing mutations within CENP-B boxes, indicating that CENP-B has some functions in assembling centromere/kinetochore components on alphoid DNA. To investigate whether any interactions exist between CENP-B and the other centromere proteins, we screened a cDNA library by yeast two-hybrid analysis. An interaction between CENP-B and CENP-C was detected, and the CENP-C domains required were determined to overlap with three Mif2 homologous regions, which were also revealed to be involved in the CENP-C assembly of centromeres by expression of truncated polypeptides in cultured cells. Overproduction of truncated CENP-B containing no CENP-C interaction domains caused abnormal duplication of CENP-C domains at G2 and cell cycle delay at metaphase. These results suggest that the interaction between CENP-B and CENP-C may be involved in the correct assembly of CENP-C on alphoid DNA. In other words, a possible molecular linkage may exist between one of the kinetochore components and human centromere DNA through CENP-B/CENP-B box interaction.
Suzuki
Nobutaka
N
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan.
Nakano
Megumi
M
Nozaki
Naohito
N
Egashira
Shin-ichiro
S
Okazaki
Tuneko
T
Masumoto
Hiroshi
H
D001324:Autoantigens; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002503:Centromere / Q000737:chemistry / Q000378:metabolism; D051936:Centromere Protein B; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D003703:Demecolcine / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D016195:G2 Phase; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000737:chemistry; D008677:Metaphase; D008858:Microscopy, Phase-Contrast; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D020798:Two-Hybrid System Techniques
D001324:Autoantigens; C497926:CENPB protein, human; D051936:Centromere Protein B; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C083768:MIF2 protein, S cerevisiae; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; C060905:centromere protein C; D004247:DNA; D003703:Demecolcine