title: 5-Methylcytosine (5mC) and 5-Hydroxymethylcytosine (5hmC) Enhance the DNA Binding of CREB1 to the C/EBP Half-Site Tetranucleotide GCAA.
abstract: In human and mouse stem cells and brain, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) can occur outside of CG dinucleotides. Using protein binding microarrays (PBMs) containing 60-mer DNA probes, we evaluated the effect of 5mC and 5hmC on one DNA strand on the double-stranded DNA binding of the mouse B-ZIP transcription factors (TFs) CREB1, ATF1, and JUND. 5mC inhibited binding of CREB1 to the canonical CRE half-site |GTCA but enhanced binding to the C/EBP half-site |GCAA. 5hmC inhibited binding of CREB1 to all 8-mers except TGAT|GCAA, where binding is enhanced. We observed similar DNA binding patterns with ATF1, a closely related B-ZIP domain. In contrast, both 5mC and 5hmC inhibited binding of JUND. These results identify new DNA sequences that are well-bound by CREB1 and ATF1 only when they contain 5mC or 5hmC. Analysis of two X-ray structures examines the consequences of 5mC and 5hmC on DNA binding by CREB and FOS|JUN.
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Khund Sayeed
authors.initials: KS
authors.lastname: Syed
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Ximiao
authors.initials: X
authors.lastname: He
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Desiree
authors.initials: D
authors.lastname: Tillo
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Stewart R
authors.initials: SR
authors.lastname: Durell
authors.affiliation: Laboratory of Metabolism and ‡Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health , Room 3128, Building 37, Bethesda, Maryland 20892, United States.
authors.forename: Charles
authors.initials: C
authors.lastname: Vinson
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D051379:Mice; D011401:Promoter Regions, Genetic
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; C492004:Creb1 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D004247:DNA