OsBP-5与OsEBP-89两蛋白之间相互作用区域的确定

Identification of the Regions Involved in Protein-Protein Interaction of OsBP-5 and OsEBP-89

OsBP 5 (MYC类转录因子 )与OsEBP 89(AP2 /EREBP类转录因子 )两个蛋白之间可以相互作用 ,并能协同调控水稻waxy基因的表达。将OsBP 5与OsEBP 89cDNA的不同限制性片段分别克隆到酵母双杂交系统的载体上 ,利用酵母双杂交的方法确定了OsEBP 89中与OsBP 5相互作用的区域位于AP2 /EREBP保守域的RAYD元件内 ;OsBP 5中与OsEBP 89相互作用的区域则有两个区段 ,它们分别位于Pro68与Val171之间和Leu2 84与Gly3 3 5之间 ,而不是位于HLH保守域内。酵母系统中的实验还表明 ,OsEBP 89的 3′端部分氨基酸在一定程度上防碍了它与OsBP

Two transcription factors, OsBP 5 and OsEBP 89, can interact synergically to regulate the expression of rice waxy gene. In this study, we dissected OsBP 5 cDNA and OsEBP 89 cDNA and identified that the RAYD element within the EREBP domain of OsEBP 89 was involved in its interaction with OsBP 5 (Fig.2), whereas the interacting regions of OsBP 5 were involved in the interaction with OsEBP 89 located in the sequences of both Pro 68 Val171 and Leu 284 Gly 335 (Fig.3), by using yeast two hybrid system (Fig.1). The presence of the c terminal of OsEBP 89 protein was found to hinder the interaction between the two proteins OsEBP 89 and OsBP 5 in the yeast two hybrid system.