Molecular basis of CONSTANS oligomerization in FLOWERING LOCUS T activation

The transcription factor CONSTANS(CO)integrates day-length information to induce the expression of florigen FLOWERING LOCUS T(FT)in Arabidopsis.We recently reported that the C-terminal CCT domain of CO forms a complex with NUCLEAR FACTOR-YB/YC to recognize multiple cis-elements in the FT promoter,and the N-terminal tandem B-box domains form a homomultimeric assembly.However,the mechanism and biological function of CO multimerization remained unclear.Here,we report that CO takes on a head-to-tail oligomeric configuration via its B-boxes to mediate FT activation in long days.The crystal structure of B-boxesCOreveals a closely connected tandem B-box fold forming a continuous head-to-tail assembly through unique CDHH zinc fingers.Mutating the key residues involved in CO oligomerization resulted in a non-functional CO,as evidenced by the inability to rescue co mutants.By contrast,a transgene encoding a human p53-derived tetrameric peptide in place of the B-boxesCOrescued co mutant,emphasizing the essential role of BboxesCO-mediated oligomerization.Furthermore,we found that the four TGTG-bearing cis-elements in FT proximal promoter are required for FT activation in long days.Our results suggest that CO forms a multimer to bind to the four TGTG motifs in the FT promoter to mediate FT activation.