Regulating RNAPII levels: gaining insight into the translation, assembly and transport of RNAPII subunits.

Informazioni relative al progetto

RNAPII biogenesis

ID dell’accordo di sovvenzione: 101066734

DOI

10.3030/101066734

Progetto chiuso

Data della firma CE 18 Luglio 2022

Data di avvio 1 Settembre 2023

Data di completamento 30 Settembre 2025

Finanziato da

• Marie Skłodowska-Curie Actions (MSCA)

Costo totale

Nessun dato

Contributo UE

€ 214 934,40

Coordinato da

KOBENHAVNS UNIVERSITET
Denmark

Periodic Reporting for period 1 - RNAPII biogenesis (Regulating RNAPII levels: gaining insight into the translation, assembly and transport of RNAPII subunits.)

Periodo di rendicontazione: 2023-09-01 al 2025-09-30

Sintesi del contesto e degli obiettivi generali del progetto

RNA polymerase II (RNAPII), the key enzyme responsible for the transcription of all protein-coding genes and many non-coding genes in eukaryotes. RNAPII is a heteromeric complex, consisting of twelve subunits (RPB1-RPB12). Emerging evidence from our lab, as well as from others, has suggested that the cellular abundance of RPB1 is tightly regulated in cells, and that it can act as a regulatory node for transcription. Nevertheless, the mechanisms involved in sensing and regulating RPB1 levels, and in extension RNAPII abundance, remained poorly understood.

The objective of the project was therefore to gain more insight into how RNAPII levels are regulated in human cells. To this end, I had proposed three work packages that focused on different steps in the RNAPII biogenesis pathway, including synthesis of the RPB1 subunit synthesis, assembly of the entire RNAPII heteromeric complex, and its transport into the nucleus.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

During the course of this action, we have achieved several important milestones. Specifically, we have identified a group of RNA-binding proteins that interact with the RPB1 mRNA and are likely involved in its post-transcriptional regulation. We are currently following up on these candidates with functional assays to gain further insight into their role in regulating RPB1 levels. In addition, we confirmed that a conserved stretch of nucleotides in the 3’UTR of RPB1 is important for the regulation of RPB1 mRNA, and have preliminary data suggesting that it might be important for the transcriptional activity of RNAPII. Moreover, we have uncovered that RNAPII subunits are assembled co-translationally in human cells, and are in the process of elucidating the precise mechanism by which this occurs.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

We have generated several rich datasets that have provided insight into the proteins that are involved in the post-transcriptional regulation of RPB1, the main subunit of RNAPII. Moreover, we have generated cell lines, which have improved upon the current tools available to study the assembly of RNAPII in vivo. These tools have been invaluable in gaining insight into the biogenesis of RNAPII and have opened up further avenues for mechanistic research that are currently being explored. These findings not only provides a better understanding of transcription, but also open up new avenues for therapeutic strategies. For example, different modes of inhibition of RNAPII-mediated transcription are currently investigated as a potential therapy for cancer.

Schematic of work packages