Isolation and characterization of the binding sequences for the product of the Arabidopsis floral homeotic gene AGAMOUS

Huang, H., Mizukami, Y., Hu, Y., Ma, H. (October 1993) Isolation and characterization of the binding sequences for the product of the Arabidopsis floral homeotic gene AGAMOUS. Nucleic Acids Res, 21 (20). pp. 4769-76. ISSN 0305-1048 (Print)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/7901838
DOI: 10.1093/nar/21.20.4769

Abstract

The Arabidopsis floral homeotic gene AGAMOUS (AG) is required for normal flower development. The deduced AG protein contains a region which shares substantial sequence similarity with the DNA-binding domains of known transcription factors, SRF (human) and MCM1 (yeast). Therefore, it is likely that AG is also a DNA-binding protein regulating transcription of floral genes. We describe here several experiments to characterize AG-DNA binding in vitro. We show that AG indeed binds a DNA sequence matching the consensus of SRF targets. Further, we have selected the AG-binding sequences from a pool of random oligonucleotides, and deduced an AG-binding consensus sequence of TT(A/T)CC(A/T)(A/t)2(T/A)NNGG(-G)(A/t)2. We have demonstrated that AG binds to the consensus region of three of the oligonucleotides by footprinting analysis. Finally, we have examined AG's relative binding affinity for different sequences, as compared to SRF, by gel mobility shift analysis. Our results indicate that AG is a sequence-specific DNA-binding protein, and that the AG-binding consensus sequence is similar to those of MCM1 and SRF.

Item Type: Paper
Uncontrolled Keywords: AGAMOUS Protein, Arabidopsis Arabidopsis/ genetics/growth & development Base Sequence Binding Sites Cloning, Molecular Consensus Sequence DNA/metabolism DNA-Binding Proteins/ genetics/metabolism Genes, Homeobox Genes, Plant Molecular Sequence Data Plant Proteins/ genetics/metabolism Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.
Subjects: organism description > plant > Arabidopsis
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > homeobox genes
organism description > plant
CSHL Authors:
Communities: CSHL labs
Depositing User: Matt Covey
Date: 11 October 1993
Date Deposited: 08 Apr 2016 16:26
Last Modified: 08 Nov 2017 16:10
PMCID: PMC331504
Related URLs:
URI: http://repository.cshl.edu/id/eprint/32581

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