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Light-dependent composition of transcriptional complexes in plants

Project description

Throwing the spotlight on transcription mechanisms in plants

Plants have developed sophisticated mechanisms to respond to environmental changes through interconnected signalling pathways. Information about transcriptional regulation in response to various stimuli in plants relies mainly on identification of the signalling pathways and transcription agents in a particular response. The EU-funded TRANSLIGHT project will use two complementary methods based on quantitative mass spectrometry to insulate protein complexes in plant cells at different phases of the transcription cycle and in response to light. It will combine these methods to identify region-specific transcription mechanisms in plants and study the dynamic composition of these complexes in response to light.

Objective

Due to their sessile nature, plants have evolved very sophisticated mechanisms to respond and adjust to the changes in environmental conditions. They do so through interconnected signaling pathways that trigger global changes in gene expression. Information about transcriptional regulation in response to different stimuli in plants relies mostly on the identification of the signaling pathways and transcription factors involved in a particular response. Importantly, investigations in yeast and humans indicate that the activity of the RNA Polymerase II (RNAPII) complex itself is subjected to a tight regulation by multiple interacting factors that respond to endogenous and exogenous stimuli. Therefore, environment-dependent changes in the activity of this transcriptional machinery seem a plausible new layer of regulation to fine-tune the transcriptional response to external stimuli in plants. In this proposal, we aim to use two complementary approaches based on quantitative mass spectrometry to isolate protein complexes in plant cells at different phases of the transcription cycle and, more important, in response to a major environmental stimulus, light. The first approach relies on the use of deactivated CRISPR/Cas9 to target proximal promoter regions and isolate proteins complexes involved in early events of light-induced transcription. In the second approach, we will identify RNAPII-associated complexes specific of transcription initiation and late elongation/termination. It relies on the use of antibodies that recognize Ser5P and Ser2P marks in the RNAPII, hallmarks of transcription initiation and termination, respectively, to isolate the region-specific RNAPII-associated complexes. Together, these two approaches will allow us i) to identify region-specific transcription machinery components in plants, both conserved and likely plant-specific ii) to study the dynamic composition of these complexes during the response to light.

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Coordinator

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Net EU contribution
€ 160 932,48
Address
Calle serrano 117
28006 Madrid
Spain

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Region
Comunidad de Madrid Comunidad de Madrid Madrid
Activity type
Research Organisations
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Other funding
€ 0,00