Floral transcription factor AGAMOUS interacts in vitro with a leucine-rich repeat and an acid phosphatase protein complex.
We are interested in identifying potential protein interactors of MADS domain transcription factors during Arabidopsis thaliana flower development. We based our biochemical search on a conserved motif in the MADS domain that includes putative phosphatase and phosphorylation sites that may mediate protein interactions. An affinity column with this motif and a few surrounding hypervariable amino acids derived from the AGAMOUS sequence was prepared and used to isolate potential interactors from floral crude extracts. Only two proteins were specifically bound to the affinity column. The first corresponds to a carpel specific storage protein, VSP1, that presents acid phosphatase activity, and the second is a novel leucine-rich repeat protein that we have named FLOR1. Coimmunoprecipitation, two-hybrid yeast, and affinity column assays show that the FLOR1-VSP1 complex interacts with AGAMOUS and that this transcription factor directly interacts with FLOR1. This is the first assay to show an interaction between plant MADS domain factors and non-MADS proteins.
Gamboa
A
A
Laboratorio de Genética Molecular y Evolución, Instituto de Ecología, Universidad Nacional Autónoma de México, Ap. Postal 70-275, México DF, 04510, México.
Paéz-Valencia
J
J
Acevedo
G F
GF
Vázquez-Moreno
L
L
Alvarez-Buylla
R E
RE
D026342:AGAMOUS Protein, Arabidopsis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000135:Acid Phosphatase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism*; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D018514:Plant Structures / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D016415:Sequence Alignment; D020798:Two-Hybrid System Techniques
D026342:AGAMOUS Protein, Arabidopsis; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; C441478:Flor1 protein, Arabidopsis; D046911:Macromolecular Substances; D008565:Membrane Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; C441479:VSP1 protein, Arabidopsis; D000135:Acid Phosphatase; D010450:Endopeptidases; D007930:Leucine