UV-B signaling pathways and fluence rate dependent transcriptional regulation of ARIADNE12.

ABSTRACT

ARI12 belongs to a family of 'RING between RING fingers' (RBR) domain proteins with E3 ligase activity (Eisenhaber et al. 2007). The Arabidopsis genome codes for 14 ARI genes and two pseudogenes (Mladek et al. 2003). Under standard growth conditions ARI12 is predominantly expressed in roots. In addition, ARI12 is strongly induced in leaves following exposure to ultraviolet (UV)-B radiation at dosages similar to those in areas under a reduced ozone layer. With quantitative reverse transcription polymerase chain reaction analyses and promoterreporter constructs we show that the expression of ARI12 peaks 2-4 h after UV-B radiation exposure. To test if ARI12's transcriptional activation depends on key players of the UV-B signaling pathway, ARI12 expression was quantified in mutants of the ELONGATED HYPOCOTYL5 (HY5), HY5 HOMOLOG (HYH) and the UV RESISTANCE LOCUS8 (UVR8) genes. ARI12 transcription was reduced by 50-70% in hy5, hyh and hy5/hyh double mutants, but not in uvr8 mutants. However, under low fluence rate UV-B conditions ARI12 is not induced in these mutants. Our results show that ARI12 represents a downstream target of the low fluence rate UVR8/HY5/HYH UV-B signaling pathway while under high fluence rates its expression is regulated by the two bZIP transcription factors HY5 and HYH in an UVR8-independent manner.