CENP-A associated complex satellite DNA in the kinetochore of the Indian muntjac

The centromere/kinetochore complex is a chromosomal assembly that mediates chromosome motility and mitotic regulation by interacting with microtubules of the mitotic spindle apparatus. Centromere protein A (CENP-A) is a histo ne H3 homolog that is concentrated in the chromatin of the inner kinetochor e plate of human chromosomes, To identify DNA sequences associated with the inner kinetochore plate, we used anticentromere autoantibodies to immunopr ecipitate CENP-A associated chromatin selectively from Indian muntjac fibro blasts. DNA was cloned from immunoprecipitated CENP-A- associated chromatin and characterized by DNA sequence and hybridization analyses. A novel cent romeric satellite DNA sequence was identified and shown by fluorescence in situ hybridization analysis to be present at all centromeres of the Indian muntjac. This satellite DNA constitutes a 972 bp monomer repeat and shows p al tial homology with satellite II DNA of the white-tailed deer. Southern b lot analysis of muntjac genomic DNA suggests that this satellite DNA is pre sent in repetitive tandem arrays and contains complex internal arrangements . In conjunction with previous work showing the association of CENP-A with human alpha-satellite DNA, we conclude that the mammalian inner kinetochore glare contains a unique form of chromatin that contains CENP-A in associat ion with complex satellite DNA.