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© 1994 Oxford University Press

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Distribution of CENP-B boxes reflected in CREST centromere antigenic sites on long-range {alpha}-satellite DNA arrays of human chromosome 21

Masashi Ikeno, Hiroshi Masumoto* and Tuneko Okazaki

Department of Molecular Biology, School of Science, Nagoya University Chikusa-ku, Nagoya 464–01, Japan

*To whom correspondence should be addressed

Received March 22, 1994; Accepted June 6, 1994

The long-range organization of alphold DNA arrays of human chromosome 21 was investigated using a mouse - human somatic cell hybrid. Two distinct long alphoid DNA arrays, the loci {alpha}21-l and {alpha}21-ll, were Identified In the centromere region of human chromosome 21. The {alpha}21-l locus, composed of an array of 11 monomer repeat units (the 11 mer), was estimated to have a total length of 1.3 Mbp. CENP-B boxes, the binding sites of the centromere protein B (CENP-B), appeared In every other monomer unit in the 11 mer except for one place where two monomer units were repeated without any CENP-B box. The other locus, {alpha}21-ll, was found to be composed of alphoid subfamilies with low homology to the components of {alpha}21-l locus. Five different alphoid clones presenting 32 monomer units In total were isolated from the {alpha}21-ll locus. Sequences of these monomer units diverged between 71 – 89% and no unit containing a CENP-B box was found. By analysis using two color FISH, the {alpha}21-l was localized to the primary constriction, whereas the {alpha}21-ll site was located slightly to the short arm side. Furthermore, a combination of FISH and immuno-fluorescent staining indicated that the {alpha}21-l site was co-localized and overlapped with the CREST centromere antigenic site on mitotic chromosomes and In interphase nuclei, while a21-ll was distributed broadly. Our data suggest that the locus a21-l containing regularly spaced CENP-B boxes at high-frequency and the assembly site of the centromere antigens may be involved in common centromere function in both human and mouse cells.


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