A human centromere antigen (CENP-B) interacts with a short specific sequence in alphoid DNA, a human centromeric satellite.
We report the interaction between a human centromere antigen and an alphoid DNA, a human centromeric satellite DNA, which consists of 170-bp repeating units. A cloned alphoid DNA fragment incubated with a HeLa cell nuclear extract is selectively immunoprecipitated by the anticentromere sera from scleroderma patients. Immunoprecipitation of the DNA made by primer extension defines the 17-bp segment on the alphoid DNA that is required for formation of DNA-antigen complex. On the other hand, when proteins bound to the biotinylated alphoid DNA carrying the 17-bp motif are recovered by streptavidin agarose and immunoblotted, the 80-kD centromere antigen (CENP-B) is detected. DNA binding experiments for proteins immunoprecipitated with anticentromere serum, separated by gel electrophoresis, and transferred to a membrane strongly suggest that the 80-kD antigen specifically binds to the DNA fragment with the 17-bp motif. The 17-bp motif is termed the "CENP-B box." Alphoid monomers with the CENP-B box are found in all the known alphoid subclasses, with varying frequencies, except the one derived from the Y chromosome so far cloned. These results imply that the interaction of the 80-kD centromere antigen with the CENP-B box in the alphoid repeats may play some crucial role in the formation of specified structure and/or function of human centromere.
Masumoto
H
H
Department of Molecular Biology, School of Science, Nagoya University, Japan.
Masukata
H
H
Muro
Y
Y
Nozaki
N
N
Okazaki
T
T
D001324:Autoantigens; D001483:Base Sequence; D002467:Cell Nucleus / Q000276:immunology; D002503:Centromere / Q000276:immunology*; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D002875:Chromosomes / Q000276:immunology*; D003001:Cloning, Molecular; D004276:DNA, Satellite / Q000276:immunology* / Q000302:isolation & purification; D004268:DNA-Binding Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids; D011485:Protein Binding; D012595:Scleroderma, Systemic / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid
D001324:Autoantigens; C497926:CENPB protein, human; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D004276:DNA, Satellite; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins