Analysis of centromere identity and structure in human cells

我们利用染色质IP、原子力显微镜测量和质谱分光光度法分析了HeLa细胞中含有CENPA的人着丝染色质。到目前为止，我们已经确定了CENPA染色质结构在间期是独特的，并且与包装基因组的其他染色质具有根本不同的特征。我们计划在整个细胞周期内使用实时成像和AFM技术来扩展我们的分析，以解决CENP-A如何组装和保留，以及它如何参与异位位点的非着丝粒功能。我们发现，在细胞周期中，CENP-A会改变其结构。该论文发表在2012年8月的《细胞》杂志上，并在《细胞》和《自然评论》中得到重点报道。后续研究发表在2013年1月的《Nucleus》杂志上，与这项工作相关的其他数据发表在《Nat. Struct》杂志上。Mol Bio， 2014年1月。我们还撰写了一本书的章节，介绍了我们实验室用于分析特定染色质结构的方法和技术（Walkiewicz et al methods Mol Bio 2014），以及对染色质生物学领域当前文献的综述(Volle和Dalal， Curr。当今。Gen. Dev, 2014)。

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).