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Electrophoretic mobility shift assay reveals a novel recognition sequence for Setaria italica NAC protein

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Title Electrophoretic mobility shift assay reveals a novel recognition sequence for Setaria italica NAC protein
 
Creator Puranik, Swati
Kumar, Karunesh
Srivastava, Prem S.
Prasad, Manoj
 
Subject Setaria italica
NAC transcription factor
EMSA
cis-element
recognition sequence
 
Description The NAC (NAM/ATAF1,2/CUC2) proteins are among the largest family of plant transcription factors. Its members have been associated with diverse plant processes and intricately regulate the expression of several genes. Inspite of this immense progress, knowledge of their DNA-binding properties are still limited. In our recent publication,1 we reported isolation of a membrane-associated NAC domain protein from Setaria italica (SiNAC). Transactivation analysis revealed that it was a functionally active transcription factor as it could stimulate expression of reporter genes in vivo. Truncations of the transmembrane region of the protein lead to its nuclear localization. Here we describe expression and purification of SiNAC DNA-binding domain. We further report identification of a novel DNA-binding site, [C/G][A/T][T/A][G/C]TC[C/G][A/T][C/G][G/C] for SiNAC by electrophoretic mobility shift assay. The SiNAC-GST protein could bind to the NAC recognition sequence in vitro as well as to sequences where some bases had been reshuffled. The results presented here contribute to our understanding of the DNA-binding specificity of SiNAC protein.
We gratefully acknowledge the financial support from the Department of
Biotechnology, Government of India (BT/
PR9851/AGR/02/521/2007).
 
Date 2014-04-30T10:34:40Z
2014-04-30T10:34:40Z
2011
8 July 2011
 
Type Article
 
Identifier Plant Signal. Behav., 6(10): 1588-1590
http://hdl.handle.net/123456789/211
 
Language en
 
Publisher Landes Bioscience