Analysis of centromere identity and structure in human cells

我们用染色质IP分析了培养的HeLa细胞中含有CENPA的人着丝粒染色质，然后用原子力显微镜测量和质量光谱分析。到目前为止，我们已经确定CENPA染色质结构是在间期唯一组织的，并且具有与包装基因组的其他染色质根本不同的特征。我们计划使用实时成像和AFM在细胞周期中扩展我们的分析，以解决CENP-A是如何组装和保留的，以及它如何参与异位基因座的非着丝粒功能。我们发现CENP-A在细胞周期中改变了它的结构。这篇手稿已于2012年8月发表在《细胞》杂志上，并在《细胞与自然评论》中重点发表。后续研究已于2013年1月发表在《核》杂志上，与这项工作相关的更多数据也发表在《NAT》杂志上。结构。2014年1月，Mol Bio。我们还写了一本书的章节，介绍了在我们实验室中用于分析特殊染色质结构的方法和技术(Walkiewicz等人的方法Mol Bio 2014)，以及对染色质生物学领域当前文献的回顾(Volle和Dalal，Curr。奥平。Dev将军，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).