Molecular Structure of the GARP Family of Plant Myb-Related DNA Binding Motifs of the Arabidopsis Response Regulators
| Autor: | Takeshi Mizuno, Aya Imamura, Toshimasa Yamazaki, Tomohisa Hatta, Kazuo Hosoda, Etsuko Katoh, Hisami Yamada, Mari Tachiki |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular Magnetic Resonance Spectroscopy Protein Conformation Recombinant Fusion Proteins Green Fluorescent Proteins Molecular Sequence Data Nuclear Localization Signals Arabidopsis Plant Science DNA-binding protein Proto-Oncogene Proteins c-myb chemistry.chemical_compound MYB Amino Acid Sequence Transcription factor Plant Proteins Genetics Binding Sites Base Sequence Sequence Homology Amino Acid biology Arabidopsis Proteins Temperature Cell Biology biology.organism_classification Fusion protein Cell biology DNA-Binding Proteins DNA binding site Luminescent Proteins chemistry Nuclear localization sequence DNA Signal Transduction Transcription Factors Research Article |
| Zdroj: | The Plant Cell. 14:2015-2029 |
| ISSN: | 1532-298X 1040-4651 |
| DOI: | 10.1105/tpc.002733 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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