Targeting of EZH2 to a defined genomic site is sufficient for recruitment of Dnmt3a but not de novo DNA methylation. Export

@article{citeulike:5720033, abstract = {Polycomb-mediated gene silencing and DNA methylation underlie many epigenetic processes important in normal development as well as in cancer. An interaction between EZH2 of the Polycomb repressive complex 2 (PRC2), which trimethylates lysine 27 on Histone 3 (H3K27me3), and all three DNA methyltransferases (DNMTs) has been reported, implicating a role for PRC2 in directing DNA methylation. Interestingly, however, the majority of H3K27me3 marked genes lack DNA methylation in ES cells, indicating that EZH2 recruitment may not be sufficient to promote DNA methylation. Here, we employed a Gal4DBD/gal4UAS-based system to directly test if EZH2 binding at a defined genomic site is sufficient to promote de novo DNA methylation in a murine erythroleukaemia cell line. Targeting of a Gal4DBD-EZH2 fusion to an intergenic transgene bearing a gal4 binding-site array promoted localized recruitment of Suz12 and Bmi1, subunits of PRC2 and PRC1, respectively, and deposition of H3K27me3. Further analysis of the H3K27me3-marked site revealed the persistence of H3K4me2, a mark inversely correlated with DNA methylation. Strikingly, while Dnmt3a was also recruited in an EZH2-dependent manner, de novo DNA methylation of the transgene was not observed. Thus, while targeting of EZH2 to a specific genomic site is sufficient for recruitment of Dnmt3a, additional events are required for de novo DNA methylation.}, author = {Rush, Margaret and Appanah, Ruth and Lee, Sandra and Lam, Lucia L. and Goyal, Preeti and Lorincz, Matthew C.}, citeulike-article-id = {5720033}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/19717977}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=19717977}, day = {29}, issn = {1559-2308}, journal = {Epigenetics : official journal of the DNA Methylation Society}, keywords = {methylation, polycomb}, month = {August}, number = {6}, posted-at = {2009-09-11 19:06:35}, priority = {4}, title = {Targeting of EZH2 to a defined genomic site is sufficient for recruitment of Dnmt3a but not de novo DNA methylation.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/19717977}, volume = {4}, year = {2009} }

TY - JOUR ID - citeulike:5720033 L3 - citeulike-article-id:5720033 N2 - Polycomb-mediated gene silencing and DNA methylation underlie many epigenetic processes important in normal development as well as in cancer. An interaction between EZH2 of the Polycomb repressive complex 2 (PRC2), which trimethylates lysine 27 on Histone 3 (H3K27me3), and all three DNA methyltransferases (DNMTs) has been reported, implicating a role for PRC2 in directing DNA methylation. Interestingly, however, the majority of H3K27me3 marked genes lack DNA methylation in ES cells, indicating that EZH2 recruitment may not be sufficient to promote DNA methylation. Here, we employed a Gal4DBD/gal4UAS-based system to directly test if EZH2 binding at a defined genomic site is sufficient to promote de novo DNA methylation in a murine erythroleukaemia cell line. Targeting of a Gal4DBD-EZH2 fusion to an intergenic transgene bearing a gal4 binding-site array promoted localized recruitment of Suz12 and Bmi1, subunits of PRC2 and PRC1, respectively, and deposition of H3K27me3. Further analysis of the H3K27me3-marked site revealed the persistence of H3K4me2, a mark inversely correlated with DNA methylation. Strikingly, while Dnmt3a was also recruited in an EZH2-dependent manner, de novo DNA methylation of the transgene was not observed. Thus, while targeting of EZH2 to a specific genomic site is sufficient for recruitment of Dnmt3a, additional events are required for de novo DNA methylation. IS - 6 JF - Epigenetics : official journal of the DNA Methylation Society SN - 1559-2308 TI - Targeting of EZH2 to a defined genomic site is sufficient for recruitment of Dnmt3a but not de novo DNA methylation. VL - 4 KW - methylation KW - polycomb AU - Rush, Margaret AU - Appanah, Ruth AU - Lee, Sandra AU - Lam, Lucia AU - Goyal, Preeti AU - Lorincz, Matthew PY - 2009/08/29/ UR - http://view.ncbi.nlm.nih.gov/pubmed/19717977 ER -