Chandler John W, Cole Melanie, Flier Annegret, Werr Wolfgang
Institute of Developmental Biology, University of Cologne, Gyrhofstrasse 17, 50923 Cologne, Germany.
Plant Mol Biol. 2009 Jan;69(1-2):57-68. doi: 10.1007/s11103-008-9405-6. Epub 2008 Oct 2.
The BIM1 protein which has been implicated in brassinosteroid (BR) signal transduction was identified from a two hybrid screen using the N-terminus, including the AP2 domain, of the transcription factors DORNROESCHEN (DRN) and DORNROESCHEN-LIKE (DRNL) which control embryonic patterning. The protein-protein interaction between BIM1 and DRN or DRNL was confirmed by co-immunoprecipitation and for DRN also in vivo by bimolecular fluorescence complementation. BIM1 can also physically interact with PHAVOLUTA (PHV), another interaction partner of DRN and DRNL. Loss of BIM1 function results in embryo patterning defects at low penetrance, including cell division defects in the hypophyseal region and apical domain defects such as cotyledon fusion and polycotyledony, in addition to polyembryony. BIM1 expression overlaps with that of DRN and DRNL from early globular embryo stages onwards. Higher order mutants between bim1, drn, drnl and phv suggest that although BIM1 may act partially redundantly with DRN in early embryo development, all genes function within the same pathway determining cotyledon development, supporting the hypothesis that they participate in a multimeric transcription factor complex. A role of BIM1 in embryonic development not only implicates a function for brassinosteroids in this process, but the interaction of BIM1 with DRN, involved with auxin signalling, represents a possible point of hormonal crosstalk in embryonic patterning and the first example of an interaction of components of the auxin and BR signalling pathways.
通过双杂交筛选,利用控制胚胎模式形成的转录因子多恩罗申(DRN)和多恩罗申样蛋白(DRNL)的N端(包括AP2结构域),鉴定出了与油菜素类固醇(BR)信号转导有关的BIM1蛋白。BIM1与DRN或DRNL之间的蛋白质-蛋白质相互作用通过免疫共沉淀得到证实,对于DRN,还通过双分子荧光互补在体内得到证实。BIM1还能与DRN和DRNL的另一个相互作用伙伴卷须蛋白(PHV)发生物理相互作用。BIM1功能缺失会导致低频率的胚胎模式形成缺陷,包括垂体区域的细胞分裂缺陷以及顶端区域缺陷,如子叶融合和多子叶现象,此外还会出现多胚现象。从早期球形胚阶段开始,BIM1的表达就与DRN和DRNL的表达重叠。BIM1、DRN、DRNL和PHV之间的高阶突变体表明,尽管BIM1在早期胚胎发育中可能与DRN部分冗余,但所有基因都在决定子叶发育的同一途径中发挥作用,支持了它们参与多聚体转录因子复合物的假说。BIM1在胚胎发育中的作用不仅意味着油菜素类固醇在这一过程中发挥功能,而且BIM1与参与生长素信号传导的DRN之间的相互作用,代表了胚胎模式形成中激素相互作用的一个可能点,也是生长素和BR信号通路成分相互作用的首个例子。