H3K36me2/3 Binding and DNA Binding of the DNA Methyltransferase DNMT3A PWWP Domain Both Contribute to its Chromatin Interaction. Issue 24 (6th December 2019)

• Record Type: Journal Article
• Authors: Dukatz, Michael; Holzer, Katharina; Choudalakis, Michel; Emperle, Max; Lungu, Cristiana; Bashtrykov, Pavel; Jeltsch, Albert
• Is Part Of: Journal of molecular biology. Volume 431:Issue 24(2019)
• Page Start: 5063
• Page End: 5074
• Publication Date: 2019-12-06
• Subjects: DNA methylation -- DNA methyltransferase -- Chromatin binding -- PWWP domain -- DNMT3A; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.09.006
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: The PWWP domain of DNMT3 DNA methyltransferases binds to histone H3 tails containing methylated K36, and this activity is important for heterochromatic targeting. Here, we show that the PWWP domain of mouse DNMT3A binds to H3K36me2 and H3K36me3 with a slight preference for H3K36me2. PWWP domains have also been reported to bind to DNA, and the close proximity of H3K36 and nucleosomal DNA suggests a combined binding to H3K36me2/3 and DNA. We show here that the DNMT3A PWWP domain binds to DNA with a weak preference for AT-rich sequences and that the designed charge reversal R362E mutation disrupts DNA binding. The K295E mutation, as well as K295I recently identified in paraganglioma, a rare neuroendocrine neoplasm, disrupts both DNA and H3K36me2/3 binding, which is in agreement with the proximity of K295 to residues involved in K36me2/3 methyllysine binding. Nucleosome pulldown experiments show that DNA binding and H3K36me2/3 binding are important for the interaction of the DNMT3A PWWP domain with nucleosomes. Localization studies of transiently transfected fluorescently-tagged wild-type and PWWP-mutated full-length DNMT3A indicate that both interactions contribute to the subnuclear localization of DNMT3A in mouse cells. In summary, our data demonstrate that the combined binding of the DNMT3A PWWP domain to the H3 tail containing K36me2/3 and to the nucleosomal or linker DNA is important for its chromatin interaction and subnuclear targeting of DNMT3A in living cells. Graphical abstract: Image 1 Highlights: The PWWP domain of DNMT3A equally binds H3K36me2 and H3K36me3. DNMT3A-PWWP binds to DNA and the R362E mutation disrupts DNA binding. Combined H3K36me2/3 and DNA binding is important for nucleosome interaction. H3K36me2/3 and DNA binding contribute to sub-nuclear targeting of DNMT3A. K295I observed in paraganglioma disrupts DNA and H3K36me2/3 binding.