domain requirements and sequence specificity of dna binding for the forkhead transcription factor foxp3域的需求和序列特异性dna结合的forkhead转录因子foxp3.pdfVIP

Domain Requirements and Sequence Specificity of DNA Binding for the Forkhead Transcription Factor FOXP3 . .¤ Kian Peng Koh , Mark S. Sundrud , Anjana Rao* Department of Pathology, Harvard Medical School and Immune Disease Institute, Boston, Massachusetts, United States of America Abstract The forkhead, winged-helix transcription factor FOXP3 is preferentially expressed in T regulatory (Treg) cells and is critical for their immunosuppressive function. Mutations that abolish FOXP3 function lead to systemic autoimmunity in mice and humans. However, the manner by which FOXP3 recognizes cognate DNA elements is unclear. Here we identify an in vitro optimized DNA sequence to assess FOXP3 DNA binding by electrophoretic mobility shift assay (EMSA). The optimized sequence contains two tandem copies of a core DNA element resembling, but not identical to, the canonical forkhead (FKH) binding element. The tandem nature of this optimized FOXP3-binding oligonucleotide suggests a requirement for multimerization, and EMSA experiments confirm that both the DNA-binding FKH domain and an intact leucine-zipper domain, which mediates homo-multimerization of FOXP3, are required for DNA binding. These results establish a practical framework for understanding the molecular basis by which FOXP3 regulates gene transcription and programs Treg suppressive function. Citation: Koh KP, Sundrud MS, Rao A (2009) Domain Requirements and Sequence Specificity of DNA Binding for the Forkhead Transcription Factor FOXP3. PLoS ONE 4(12): e8109. doi:10.1371/journal.pone.0008109 Editor: Derya Unutmaz, New York University School of Medicine, United States of America Received October 19, 2009; Accepted November 3, 2009; Publ