Solution structure of the major DNA-binding domain of Arabidopsis thaliana ethylene-insensitive3-like3.
Ethylene-insensitive3 (EIN3) and EIN3-like (EIL) proteins are essential transcription factors in the ethylene signaling of higher plants. The EIN3/EIL proteins bind to the promoter regions of the downstream genes and regulate their expression. The location of the DNA-binding domain (DBD) in the primary structure was unclear, since the proteins show no sequence similarity to other known DBDs. Here, we identify the major DBD of an EIN3/EIL protein, Arabidopsis thaliana EIL3, containing a key mutational site for DNA binding and signaling (ein3-3 site), and determine its solution structure by NMR spectroscopy. The structure consists of five alpha-helices, possessing a novel fold dissimilar to known DBD structures. By a chemical-shift perturbation analysis, a region including the ein3-3 site is suggested to be involved in DNA binding.
Yamasaki
Kazuhiko
K
Age Dimension Research Center, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566, Japan. k-yamasaki@aist.go.jp
Kigawa
Takanori
T
Inoue
Makoto
M
Yamasaki
Tomoko
T
Yabuki
Takashi
T
Aoki
Masaaki
M
Seki
Eiko
E
Matsuda
Takayoshi
T
Tomo
Yasuko
Y
Terada
Takaho
T
Shirouzu
Mikako
M
Tanaka
Akiko
A
Seki
Motoaki
M
Shinozaki
Kazuo
K
Yokoyama
Shigeyuki
S
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D020349:Surface Plasmon Resonance; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; C106987:EIN3 protein, Arabidopsis; D009687:Nuclear Proteins; D014157:Transcription Factors; D004247:DNA; D011392:Proline