Plants, as sessile organisms, need to sense and adapt to heterogeneous environments and have developed sophisticated responses by changing their cellular physiology, gene regulation, and genome stability. Recent work demonstrated heritable stress effects on the control of genome stability in plants—a phenomenon that was suggested to be of epigenetic nature. Here, we show that temperature and UV-B stress cause immediate and heritable changes in the epigenetic control of a silent reporter gene in Arabidopsis. This stress-mediated release of gene silencing correlated with pronounced alterations in histone occupancy and in histone H3 acetylation but did not involve adjustments in DNA methylation. We observed transmission of stress effects on reporter gene silencing to non-stressed progeny, but this effect was restricted to areas consisting of a small number of cells and limited to a few non-stressed progeny generations. Furthermore, stress-induced release of gene silencing was antagonized and reset during seed aging. The transient nature of this phenomenon highlights the ability of plants to restrict stress-induced relaxation of epigenetic control mechanisms, which likely contributes to safeguarding genome integrity.

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 promoter:reporter 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.

The UV-B inducible ARIADNE12 (ARI12) gene of Arabidopsis thaliana is a member of the RING-between-RING (RBR) family of E3 ubiquitin ligases for which a novel ubiquitination mechanism was identified in mammalian homologs. This RING-HECT hybrid mechanism needs a conserved cysteine which is replaced by serine in ARI12 and might affect the E3 ubiquitin ligase activity. We have shown that under photomorphogenic UV-B, ARI12 is a downstream target of the classical ultraviolet B (UV-B) UV RESISTANCE LOCUS 8 (UVR8) pathway. However, under high fluence rate of UV-B ARI12 was induced independently of UVR8 and the UV-A/blue light and red/far-red photoreceptors. A key component of several light signaling pathways is CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1). Upon UV-B COP1 is trapped in the nucleus through interaction with UVR8 permitting the activation of genes that regulate the biosynthesis of UV-B protective metabolites and growth adaptations. To clarify the role of COP1 in the regulation of ARI12 mRNA expression and ARI12 protein stability, localization and interaction with COP1 was assessed with and without UV-B. We found that COP1 controls ARI12 in white light, low and high fluence rate of UV-B. Furthermore we show that ARI12 is indeed an E3 ubiquitin ligase which is mono-ubiquitinated, a prerequisite for the RING-HECT hybrid mechanism. Finally, genetic analyses with transgenes expressing a genomic pmARI12:ARI12-GFP construct confirm the epistatic interaction between COP1 and ARI12 in growth responses to high fluence rate UV-B.