Analysis of centromere identity and structure in human cells

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We have analyzed human centromeric chromatin containing CENPA from HeLa cells in culture using Chromatin IP followed by atomic force microscopy measurements and mass spectrophotometry. Thus far, we have identified that the CENPA chromatin structure is uniquely organized during interphase, and has fundamentally different characteristics from the rest of the chromatin which packages the genome. We plan to extend our analysis using live imaging coupled with AFM across the cell cycle to address how CENP-A is assembled and retained, as well as how it participates in non-centromeric functions at ectopic loci. We have discovered that CENP-A changes its structure over the cell cycle. This manuscript has been published in the journal Cell in August 2012 and was highlighted in Cell and Nature Reviews. A follow up has been published in the journal Nucleus in January 2013, and additional data related to this work was published in Nat. Struct. Mol Bio in January 2014. We have also written a book chapter on methods and techniques used in our laboratory to analyze specialized chromatin structures (Walkiewicz et al Methods Mol Bio 2014), as well as a review of current literature in the chromatin biology field (Volle and Dalal, Curr. Opin. Gen. Dev, 2014).