REL Domain of NFATc2 Binding to Five Types of DNA Using Protein Binding Microarrays

Abstract

NFATc2 is a DNA binding protein in the Rel family transcription factors, which binds a CGGAA motif better when both cytosines in the CG dinucleotide are methylated. Using protein binding microarrays (PBMs), we examined the DNA binding of NFATc2 to three additional types of DNA: single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) with either 5-methylcytosine (5mC, M) or 5-hydroxymethylcytosine (5hmC, H) in one strand and a cytosine in the second strand. ATTTCCAC, the complement of the core GGAA motif, is better bound as ssDNA compared to dsDNA. dsDNA containing the 5-mer CGGAA with either 5mC or 5hmC in one DNA strand is bound stronger than CGGAA. In contrast, the reverse complement TTCCG is bound weaker when it contains 5mC. Analysis of the available NFATc2:dsDNA complexes rationalizes these PBM data.

Rapid profiling of transcription factor-cofactor interaction networks reveals principles of epigenetic regulation

“…NFAT family member motifs appear to have a relatively equal preference for A and G at this position and previous studies have noted NFATC2 binding to both of these types of sequences, albeit with varying affinities. 60 Strikingly, in SUDHL4 cells BRD4 and TBL1XR1 are also recruited to NFAT motifs with the same preference for adenine at position 8, suggesting that is a cell-type specific difference (Figure S1B). COF-specific differences within individual cell types were also observed.…”