PMID:12215502

Citation	Hosoda, K, Imamura, A, Katoh, E, Hatta, T, Tachiki, M, Yamada, H, Mizuno, T and Yamazaki, T (2002) Molecular structure of the GARP family of plant Myb-related DNA binding motifs of the Arabidopsis response regulators. Plant Cell 14:2015-29
Abstract	The B motif is a signature of type-B response regulators (ARRs) involved in His-to-Asp phosphorelay signal transduction systems in Arabidopsis. Homologous motifs occur widely in the GARP family of plant transcription factors. To gain general insight into the structure and function of B motifs (or GARP motifs), we characterized the B motif derived from a representative ARR, ARR10, which led to a number of intriguing findings. First, the B motif of ARR10 (named ARR10-B and extending from Thr-179 to Ser-242) possesses a nuclear localization signal, as indicated by the intracellular localization of a green fluorescent protein-ARR10-B fusion protein in onion epidermal cells. Second, the purified ARR10-B molecule binds specifically in vitro to DNA with the core sequence AGATT. This was demonstrated by several in vitro approaches, including PCR-assisted DNA binding site selection, gel retardation assays, and surface plasmon resonance analysis. Finally, the three-dimensional structure of ARR10-B in solution was determined by NMR spectroscopy, showing that it contains a helix-turn-helix structure. Furthermore, the mode of interaction between ARR10-B and the target DNA was assessed extensively by NMR spectroscopy. Together, these results lead us to propose that the mechanism of DNA recognition by ARR10-B is essentially the same as that of homeodomains. We conclude that the B motif is a multifunctional domain responsible for both nuclear localization and DNA binding and suggest that these insights could be applicable generally to the large GARP family of plant transcription factors.
Links	PubMed PMC150752
Keywords	Amino Acid Sequence; Arabidopsis/genetics; Arabidopsis/metabolism; Arabidopsis Proteins/chemistry; Arabidopsis Proteins/genetics; Arabidopsis Proteins/metabolism; Base Sequence; Binding Sites/genetics; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Green Fluorescent Proteins; Luminescent Proteins/genetics; Luminescent Proteins/metabolism; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Nuclear Localization Signals/genetics; Plant Proteins/chemistry; Plant Proteins/genetics; Plant Proteins/metabolism; Protein Conformation; Proto-Oncogene Proteins c-myb/chemistry; Proto-Oncogene Proteins c-myb/genetics; Proto-Oncogene Proteins c-myb/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Sequence Homology, Amino Acid; Signal Transduction/genetics; Signal Transduction/physiology; Temperature; Transcription Factors/genetics; Transcription Factors/physiology
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