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

Sreejana Ray; Desiree Tillo; Stewart R Durell; Syed Khund-Sayeed; Charles Vinson

doi:10.1021/acsomega.0c04069

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

ACS Omega. 2021 Feb 2;6(6):4147-4154. doi: 10.1021/acsomega.0c04069. eCollection 2021 Feb 16.

Authors

Sreejana Ray¹, Desiree Tillo¹, Stewart R Durell², Syed Khund-Sayeed¹, Charles Vinson¹

Affiliations

¹ Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Building 37, Room 5000, Bethesda, Maryland 20892, United States.
² Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Building 37, Room 5000, Bethesda, Maryland 20892, United States.

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.