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H2020Individual fellowship2016–2018

SUMO-ZNHIT6-snoRNPs · ZNHIT6: a new SUMO E3 ligase regulating RNA metabolism

Horizon 2020 — Marie Skłodowska-Curie Actions

Duration
2016-10-01 → 2018-09-30
EU contribution
€195,455
Participants
1
Scheme
MSCA-IF

Lines connect the coordinator with its partners.

Results in brief

ZNHIT6: a new SUMO E3 ligase regulating RNA metabolism

This project is focused on answering a basic biological question by performing a series of well controlled and designed experiments. My host laboratory is interested in the role of a small ubiquitin-like modifier (SUMO) in various cellular processes, which can be studied by a combination of biochemical, biological and global proteomic approaches. The overall objective of the project was to investigate the role of ZNHIT6, a putative SUMO E3 ligase in the regulation of RNA metabolism. In particular we wanted to investigate a role for this protein in the regulation of small nucleolar RNA-protein complexes (snoRNPs) that mediate the modification of rRNA nucleotides and processing of rRNA precursors. It has been previously established that SUMO-mediated protein modifications play a crucial role in the regulation of snoRNP functions and this project would therefore expand our current understanding of this important biological process. It has a potentially wider benefit to society as snoRNAs have been shown to play a role in cancer and neurodegenerative and viral diseases. During the fellowship, we have widened our research interest to also determine the biological role of ZNHIT6 in stem cell differentiation, due to its importance in general translation regulation. That might have numerous implications in regenerative medicine, cancer treatments, and our understanding of the role of ZNHIT6 and the wider SUMO network in early human development.

Data: CORDIS, © European Union

Project objective

It is widely accepted that eukaryotic cells contain large populations of small nucleolar RNA-protein complexes (snoRNPs), which mediate the modification of rRNA nucleotides and facilitate cleavage of rRNA precursors. Thus, they may control general protein translation by regulating the level and function of ribosomes. It is currently thought that understanding the process of snoRNP biogenesis will provide the knowledge indispensable to determine their exact function and means of regulation. Although, it is known that SUMO-mediated protein modifications are crucial for snoRNP functions, the E3 responsible for this process as well as their precise targets, or time and space frame of the modification are not known. Preliminary results of the host group obtained by novel proteomic approaches demonstrated that proteins of box C/D snoRNP complex undergo SUMOylation and that BCD1/ZNHIT6 might be a new SUMO E3 ligase responsible for these modifications.The specific aim of this study is to demonstrate whether ZNHIT6 indeed is a SUMO E3 ligase, to identify its substrates and reveal the function of BCD1-mediated SUMOylation of these targets in the regulation of RNA metabolism. For that purpose, I will be trained in a number of state-of the-art techniques. Notably, I will create a series of cell lines depleted of BCD1 or expressing its tagged versions using CRISPR/Cas technology. The role of ZNHIT6 in RNA metabolism will be analysed by classical biological methods, such as: IF, WB, FACS. In addition, I will use new mass spectrometry based approach designed by the host lab for the detection of SUMOylated targets. A wide range of in vitro assays will also be used to determine interactions and SUMO E3 ligase activity of ZNHIT6. The innovative character of this proposal lies in the complementarity of research techniques used and the participants involved for studying this biological question and will provide insights into the cross talk between SUMOylation and RNA metabolism.

Original text from CORDIS.

Participants

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Data: CORDIS, © European Union