文献类型：期刊文献

英文题名：Monitoring Histone Methylation (H3K9me3) Changes in Live Cells

作者：Sanchez, Oscar F.[1];Mendonca, Agnes[1];Min, Alan[2];Liu, Jichang[3];Yuan, Chongli[1,4]

机构：[1]Purdue Univ, Davidson Sch Chem Engn, 480 Stadium Mall Dr, W Lafayette, IN 47907 USA;[2]Purdue Univ, Dept Comp Sci, 480 Stadium Mall Dr, W Lafayette, IN 47907 USA;[3]East China Univ Sci & Technol, State Key Lab Chem Engn, Shanghai 200237, Peoples R China;[4]Purdue Univ, Ctr Canc Res, W Lafayette, IN 47907 USA

年份：2019

卷号：4

期号：8

起止页码：13250

外文期刊名：ACS OMEGA

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000483407600029)】；

基金：O.S. and A.M. would like to thank Dr. James A. Schaber from the Bindley Bioscience Imaging Facility for helping with cell imaging. This work was supported by US Army Medical Research [W81XWH-14-1-0012], National Science Foundation [CBET-1512285 & CBET-1705560], and Indiana Clinical and Translational Science Institute. Administrative Department of Science, Technology and Innovation from Colombia and dFulbright Colombia [529 to O.F.S]. The authors gratefully acknowledge the support from the Purdue University Center for Cancer Research, NIH grant P30 CA023168.

语种：英文

摘要：H3K9me3 (methylation of lysine 9 of histone H3) is an epigenetic modification that acts as a repressor mark. Several diseases, including cancers and neurological disorders, have been associated with aberrant changes in H3K9me3 levels. Different tools have been developed to enable detection and quantification of H3K9me3 levels in cells. Most techniques, however, lack live cell compatibility. To address this concern, we have engineered recombinant protein sensors for probing H3K9me3 in situ. A heterodimeric sensor containing a chromodomain and chromo shadow domain from HP1a was found to be optimal in recognizing H3K9me3 and exhibited similar spatial resolution to commercial antibodies. Our sensor offers similar quantitative accuracy in characterizing changes in H3K9me3 compared to antibodies but claims single cell resolution. The sensor was applied to evaluate changes in H3K9me3 responding to environmental chemical atrazine (ATZ). ATZ was found to result in significant reductions in H3K9me3 levels after 24 h of exposure. Its impact on the distribution of H3K9me3 among cell populations was also assessed and found to be distinctive. We foresee the application of our sensors in multiple toxicity and drug-screening applications.