Further evidence that CENP-C is a necessary component of active centromeres: studies of a dic(X; 15) with simultaneous immunofluorescence and FISH.

The stability of certain dicentric chromosomes in humans seems to result from inactivation of one centromere, yielding a functionally monocentric chromosome. Centromere protein C (CENP-C) was previously shown to be present at active centromeres but absent from the inactive centromere of one homologous dicentric rearrangement. We have combined indirect immunofluorescence detection of CENP-C and fluorescence in situ hybridization with chromosome-specific alpha-satellite DNA probes in a simultaneous assay to unequivocally identify the active and inactive centromeres of a dicentric (X;15) translocation. In both fibroblast and lymphoblast cell lines containing the translocation, the X chromosome centromere consistently had a primary constriction and CENP-C immunofluorescence, and is therefore the active centromere. CENP-C was never detected at the chromosome 15 centromere, which appears to be inactive. The inactivation pattern is apparently stable and observed in all cells with the translocation. Immunofluorescence with CREST serum revealed staining at both centromeres of the translocation, and thus was not specific to the active centromere. This study demonstrates the specificity of CENP-C to the active centromere in a non-homologous rearrangement and further establishes CENP-C as an essential component of a functional human centromere.