A critical arginine residue mediates cooperativity in the contact interface between transcription factors NFAT and AP-1

• DOI: 10.1073/pnas.93.24.13671
• Publication Date: 2002-07-26T14:32:33Z
• Authors: Blake R. Peterson; Li Jing Sun; Gregory L. Verdine
• Keywords: NFAT; Cooperativity; Cooperative binding; Transcription; Transcription preinitiation complex

The heterologous transcription factors NFAT and AP-1 coordinately regulate cytokine gene expression through cooperative binding to precisely juxtaposed DNA recognition elements. molecular origins of cooperativity in the are poorly understood. Herein we have used yeast one-hybrid screening alanine-scanning mutagenesis identify residues that affect interactions with on DNA. Mutation a single conserved Arg residue Ala cJun spacer region (R285A) led virtually complete abolition NFAT. DNA-binding activity alone was unaffected by R285A mutation, thus indicating this influences only. Ala-scanning mutations elsewhere AP-1, including cFos subunit, revealed no other strongly interacting positions. We conclude contacts making an especially important contact R285, these drive formation NFAT/AP-1/DNA complex. These results provide general strategy for selectively ablating between without affecting their ability act yield insights into structural basis coordinate regulation expression.