Relating Centromeric Topography in Fixed Human Chromosomes to α-Satellite DNA and CENP-B DistributionKhan W.A.a, e · Chisholm R.g · Tadayyon S.c, e · Subasinghe A.d, e · Norton P.c, e · Samarabandu J.d, e · Johnston L.J.g · Knoll J.H.a,e,f · Rogan P.K.b,e,f
Departments of aPathology and bBiochemistry, Schulich School of Medicine and Dentistry, cDepartment of Chemistry, Interface Science Western, and dImage Recognition and Intelligent Systems Laboratory, Electrical and Computer Engineering, at eWestern University, and fCytognomix Inc., London, Ont., and gSteacie Institute for Molecular Sciences, National Research Council of Canada, Ottawa, Ont., Canada
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Despite extensive analyses on the centromere and its associated proteins, detailed studies of centromeric DNA structure have provided limited information about its topography in condensed chromatin. We have developed a method with correlative fluorescence light microscopy and atomic force microscopy that investigates the physical and structural organization of α-satellite DNA sequences in the context of its associated protein, CENP-B, on human metaphase chromosome topography. Comparison of centromeric DNA and protein distribution patterns in fixed homologous chromosomes indicates that CENP-B and α-satellite DNA are distributed distinctly from one another and relative to observed centromeric ridge topography. Our approach facilitates correlated studies of multiple chromatin components comprising higher-order structures of human metaphase chromosomes.
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