Resumen de New mechanisms of della protein regulation and activity in arabidopsis
DELLA proteins are plant-specific transcriptional regulators known to relay environmental information to the transcriptional networks to modulate growth and development accordingly. The view of DELLAs as signalling hubs is justified by two characteristics: first, they control the activity of a large number of transcriptional factors (TFs) and other transcriptional regulators through physical interaction; and, second, they are degraded by the 26S proteasome in response to the phytohormone gibberellin (GA), whose metabolism is very sensitive to environmental stimuli (e.g. light, temperature, salt stress). However, at least two observations indicate that this mechanistic framework is still incomplete: (i) warm temperature destabilizes the GA-insensitive DELLA rga-¿17, indicating that DELLAs cannot be destabilized only by changes in GA levels; and (ii) when found at the chromatin, DELLAs are localized not only in gene promoters, but also in gene bodies, suggesting that DELLAs may regulate transcription through interactions with proteins other than TFs. In this Thesis, we provide evidence that shows how a different E3 ubiquitin ligase controls the stability of DELLAs in a GA-independent manner, and how DELLAs regulate gene expression by directly interacting with the basal transcriptional machinery. In the first chapter, using a combination of genetic, physiological, and molecular approaches we demonstrate that DELLAs are targeted to proteolytic degradation by the E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1). We show that COP1 interacts with the DELLAs GAI and RGA in vitro and in vivo, and that it promotes their polyubiquitination. We propose that COP1 represents a major pathway to degrade DELLAs in response to shade or to warm temperature. In the second chapter, we describe the interaction between DELLAs and the transcription elongation complex Polymerase-Associated Factor 1 (Paf1c). We show that Paf1c is required for the genome-wide deposition of monoubiquitinated H2B (H2Bub), a mark necessary for the progression of RNA polymerase II (RNAPII), and that this function is largely dependent on the presence of DELLAs. Likewise, impaired Paf1c or DELLA function results in a similar alteration in the accumulation and distribution of RNAPII in the Paf1c-target genes. We propose that DELLAs would exert this action by modulating the recruitment of Paf1c to the chromatin.
These two new mechanisms underscore the importance of DELLAs as a central node in the environmental signalling network and should be considered in any potential application of DELLAs as biotechnological targets.
