The tandem complex of BEL and KNOX partners is required for transcriptional repression of ga20ox1

Two interacting (t) under bar hree (a) under bar mino acid (l) under bar oop (e) under bar xtension (TALE) proteins of potato, StBEL5 (Solanum tuberosom BEL 5) and POTH1 (potato homeobox 1), mediate developmental processes by regulating phytohormone levels. Overexpression of either partner alone increased tuber yields by lowering gibberellin (GA) levels and increasing cytokinins. Gel shift assays demonstrated that StBEL5 and POTH1 bind to the regulatory region of ga20 oxidase1 (ga20ox1) from potato, a gene encoding a key enzyme in the GA biosynthetic pathway. In tandem, StBEL5 and POTH1 had a greater binding affinity for the ga20ox1 promoter than either protein alone. The StBEL5-POTH1 heterodimer bound specifically to a composite 10-bp sequence, containing two TGAC cores. Using a transcription assay, StBEL5 and POTH1 suppressed the activity of the ga20ox1 promoter by more than 50%. Dominant negative constructs containing the protein-binding domains of StBEL5 or POTH1 blocked the repression activity of StBEL5 or POTH1, respectively. The mutated ga20ox1 promoter that could be bound by the StBEL5 or POTH1 proteins individually but not by the StBEL5-POTH1 heterodimer also abolished the repression activity of StBEL5, POTH1, and the StBEL5-POTH1 heterodimer. These results indicate that the tandem interaction of StBEL5 and POTH1 is essential for regulation of the expression of their target gene.