Isolation of putative plant transcriptional coactivators using a modified two-hybrid system incorporating a GFP reporter gene.
Dual hybrid interacting screening in yeast led to the identification of two proteins from Arabidopsis both exhibiting sequence similarity to a family of transcriptional coactivators from a diverse range of organisms. Their discovery constitutes the first description of such plant proteins. A modified yeast two-hybrid approach utilising the green fluorescent protein (GFP) of Aequora victoria was developed and used to clone one of the putative plant transcriptional coactivators from an Arabidopsis cDNA library. The two proteins, designated KIWI and KELP, can associate both hetero- and homomerically and their genes were cloned and mapped on the Arabidopsis genome. Both proteins are believed to play a role in gene activation during pathogen defence and plant development. The involvement of these proteins in general plant transcription as well as the advantages of using GFP as a reporter gene for detecting protein-protein interactions are discussed.
Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Cologne, Germany.
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Cormack
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Hahlbrock
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D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000235:genetics* / Q000302:isolation & purification*
D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D018744:DNA, Plant; C113673:KELP protein, Arabidopsis; C113674:KIWI protein, Arabidopsis; D008164:Luminescent Proteins; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D015534:Trans-Activators; D049452:Green Fluorescent Proteins