Isolation and characterization of the binding sequences for the product of the Arabidopsis floral homeotic geneAGAMOUS
Year: 1993
Type: article
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.
Source: Nucleic Acids Research
Authors: Hai Huang, Yukiko Mizukami, Yi Hu, Hong Ma
Institution: Cold Spring Harbor Laboratory
Language: English
Subfield: Plant Science
Domain: Life Sciences
Open Access status: bronze
Funder: National Science Foundation
OpenAlex ID: W2038813658