Centromere size and position in Candida albicans are evolutionarily conserved independent of DNA sequence heterogeneity

The centromere regions (CEN) of all eight chromosomes in Candida albicans have been characterized in terms of nucleotide sequence and size. The boundaries of each of the eight CEN DNA regions were mapped by chromatin immunoprecipitation-PCR using polyclonal rabbit antibodies generated against C. albicans centromere-specific protein CaCse4p (CENP-A homolog). A single 3-4.5 kb unique DNA sequence on each chromosome was found to be bound to CaCse4p. Sequence analysis revealed that the eight CEN regions in C. albicans lack any conserved DNA sequence motifs common to the group; all are quite different in overall DNA sequence. In contrast to centromeres in many organisms, the C. albicans centromeres are generally free of repeated DNA elements and transposons. However, a few small inverted repeats and long terminal repeats do occur in the centromeric and pericentric regions on a few chromosomes. We also characterized the CEN DNAs in four groups of phylogenetically divergent C. albicans strains, estimated to be separated from each other by 1-3 million years. The same eight different and unique 3-4.5 kb DNA sequences are utilized as centromeres in all of these strains. The chromosomal locations and the sizes of CEN DNAs have remained conserved, in agreement with the idea that CEN function in C. albicans is templated by heritable epigenetic mechanisms.