Multiple protein regions contribute to differential activities of YABBY proteins in reproductive development

Members of the YABBY family of putative transcription factors participate in abaxial-adaxial identity determination in lateral organs in Arabidopsis (Arabidopsis thaliana). Two YABBY genes specifically expressed in reproductive structures, CRABS CLAW (CRC) and INNER NO OUTER (INO), have additional activities, with CRC promoting nectary development and carpel fusion, and INO responding to spatial regulation by SUPERMAN during ovule development. All YABBY coding regions, except YABBY5, were able to restore outer integument growth in ino-1 mutants when expressed from the INO promoter (PRO(INO)). However, INO was the only YABBY family member that responded correctly to SUPERMAN to maintain the wild-type gynoapical-gynobasal asymmetry of the outer integument. By contrast, INO, FILAMENTOUS FLOWER, and YABBY3 failed to complement crc-1 when expressed from PRO(CRC). Roles of individual regions of CRC and INO in these effects were assessed using chimeric proteins with PRO(INO) and PRO(CRC) and the relatively constitutive cauliflower mosaic virus PRO(35S). Regions of CRC were found to contribute additively to CRC-specific functions in nectary and carpel formation, with a nearly direct relationship between the amount of CRC included and the degree of complementation of crc-1. When combined with INO sequences, the central and carboxyl-terminal regions of CRC were individually sufficient to overcome inhibitory effects of SUPERMAN within the outer integument. Reproductive phenotypes resulting from constitutive expression were dependent on the nature of the central region with some contributions from the amino terminus. Thus, the YABBY family members have both unique and common functional capacities, and residues involved in differential activities are distributed throughout the protein sequences.