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© 1995 Oxford University Press

OTHER

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

Scott L. Page, William C. Earnshaw1, K.H.Andy Choo2 and Lisa G. Shaffer*,

Department of Molecular and Human Genetics, Baylor College of Medicine Houston, TX 77030 1Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine Baltimore, MD 21205, USA 2Murdoch Institute for Research into Birth Defects, Royal Children's Hospital Parkville, Victoria 3052, Australia

*To whom correspondence should be addressed

Received October 12, 1994; Revised December 1, 1994; Accepted December 1, 1994

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 chromo-some-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.


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