Markel Heike, Chandler John, Werr Wolfgang
Institut für Entwicklungsbiologie Universität zu Köln, 50923 Köln, Germany.
Nucleic Acids Res. 2002 Nov 1;30(21):4709-19. doi: 10.1093/nar/gkf591.
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.
有证据表明，通过使用果蝇engrailed（en）基因的阻遏结构域，植物转录因子可以有效地重编程为显性负性功能。en（298）N端与SHOOTMERISTEMLESS、APETALA3、PISTILLATA和KNAT1转录因子的完整编码区之间的翻译融合体的异位表达导致反式显性功能，其表型模拟功能丧失突变体。显性负性en（298）-STM功能与糖皮质激素受体的激素结合结构域的结合提供了强有力的证据，表明表型模拟依赖于嵌合蛋白进入核区室。通过这种显性负性方法，迅速确定KNAT1由BREVIPEDICELLUS基因座编码。显性负性嵌合蛋白可能广泛用于阐明植物转录激活因子的生物学功能，并且可能适用于研究植物中的蛋白质-蛋白质相互作用。
