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

Hai Huang, Yukiko Mizukami, Yi Hu, Hong Ma

Research output: Contribution to journalArticle

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Abstract

The Arabidopsis floral homeotic geneAGAMOUS (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 AGbinding 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.

Original languageEnglish (US)
Pages (from-to)4769-4776
Number of pages8
JournalNucleic acids research
Volume21
Issue number20
DOIs
StatePublished - Oct 11 1993

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Homeobox Genes
Consensus Sequence
DNA-Binding Proteins
Minichromosome Maintenance 1 Protein
Arabidopsis
Oligonucleotides
Serum Response Factor
DNA
Electrophoretic Mobility Shift Assay
Protein Binding
Yeasts
Genes
Proteins
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

@article{f0e1a08199c04465b477d2c00cf7a13f,
title = "Isolation and characterization of the binding sequences for the product of the arabidopsis floral homeotic gene AGAMOUS",
abstract = "The Arabidopsis floral homeotic geneAGAMOUS (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 AGbinding 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.",
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Isolation and characterization of the binding sequences for the product of the arabidopsis floral homeotic gene AGAMOUS. / Huang, Hai; Mizukami, Yukiko; Hu, Yi; Ma, Hong.

In: Nucleic acids research, Vol. 21, No. 20, 11.10.1993, p. 4769-4776.

Research output: Contribution to journalArticle

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