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Biochemical analysis of the interaction between Arabidopsis thaliana AtMYB30 transcription factor and its DNA specific target site

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Abstract

AtMYB30 is well known as a transcription factor involved in cell death processes during the hypersensitive response against biotic stress in Arabidopsis thaliana. Despite the relevant function played by R2R3-MYB transcription factors in the regulation of plant gene expression, limited information exists about how these proteins interact with their DNA targets, their specificity, and binding affinity. Our findings confirm the binding of a specific DNA sequence (5′-AAACCAA) to AtMYB30, binding affinities were calculated, and the effect of DNA on AtMYB30 secondary structure was evaluated. Our data shed new light on the interaction between DNA and this important member of a plant-specific transcriptional regulator family.

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Abbreviations

CD:

Circular dichroism

DBD:

DNA-binding domain

EDTA:

Ethylenediaminetetraaceticacid

EMSA:

Electrophoretic mobility shift assay

EtOH:

Ethanol

HTH:

Helix-turn-helix

ITC:

Isothermal titration calorimetry

KD :

Equilibrium dissociation constant

NO:

Nitric oxide

PAGE:

Polyacrylamide gel electrophoresis

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Acknowledgments

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa Científica de Tecnológica do Estado de Santa Catarina (FAPESC), Financiadora de Estudos e Projetos (FINEP), and Instituto Nacional de Biologia Estrutural e Bioimagem (INBEB). We acknowledge the technical staff of CEBIME for assistance.

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  1. Centro de Biologia Molecular Estrutural - INBEB, Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Florianópolis, Brazil

    Carolina Pereira Tavares & Hernán Terenzi

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  1. Carolina Pereira Tavares
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  2. Hernán Terenzi
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Correspondence to Hernán Terenzi.

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Tavares, C.P., Terenzi, H. Biochemical analysis of the interaction between Arabidopsis thaliana AtMYB30 transcription factor and its DNA specific target site. J. Plant Biochem. Biotechnol. 25, 97–103 (2016). https://doi.org/10.1007/s13562-015-0315-9

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  • DOI: https://doi.org/10.1007/s13562-015-0315-9

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