Translational fusions with the engrailed repressor domain efficiently convert plant transcription factors into dominant-negative functions.
Evidence is provided that plant transcription factors can be efficiently reprogrammed to dominant- negative functions by the use of a repressor domain of the engrailed (en) gene from Drosophila. Ectopic expression of translational fusions between the en(298) N-terminus and the complete coding regions of the SHOOTMERISTEMLESS, APETALA3, PISTILLATA and KNAT1 transcription factors results in trans-dominant functions which phenocopy loss-of-function mutants. The combination of the dominant-negative en(298)-STM function with the hormone-binding domain of the glucocorticoid receptor provides strong evidence that phenocopies rely on the incorporation of the chimeric protein into the nuclear compartment. By this dominant-negative approach KNAT1 was rapidly identified to be encoded by the BREVIPEDICELLUS locus. Dominant-negative chimeric proteins may be of wide use to elucidate biological functions of plant transcriptional activators and may be suitable to study protein-protein interactions in planta.
Markel
Heike
H
Institut für Entwicklungsbiologie Universität zu Köln, 50923 Köln, Germany.
Chandler
John
J
Werr
Wolfgang
W
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005799:Genes, Dominant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D030821:Plants, Genetically Modified; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion / Q000235:genetics; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D029681:Arabidopsis Proteins; D029721:Drosophila Proteins; C063459:En protein, Drosophila; D018398:Homeodomain Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors