Deciphering the mRNP at premature termination codons
Gabriele Neu-Yilik
Heidelberg University Medical Center for Children and Adolescents
and Molecular Medicine Partnership Unit
Medical Faculty Heidelberg/EMBL
E-mail: gabriele.neu-yilik(at)med.uni-heidelberg.de
Andreas Kulozik
Heidelberg University Medical Center for Children and Adolescents
and Molecular Medicine Partnership Unit
Medical Faculty Heidelberg/EMBL
Email: andreas.kulozik(at)med.uni-heidelberg.de
Matthias Hentze
European Molecular Biology Laboratory (EMBL)
and Molecular Medicine Partnership Unit
Medical Faculty Heidelberg/EMBL
E-mail: hentze(at)embl.org
For more information and contact visit the MMPU blood diseases group and Group mRNA metabolism - KiTZ.
Abstract
Nonsense-mediated mRNA decay (NMD) represents one of the core quality control mechanisms of gene expression, which eliminates faulty mRNAs encoding C-terminally truncated proteins. Extending beyond the basic biologic significance, NMD is also medically highly relevant because it represents an important modifier of inherited and aquired diseases. Therefore, it is important to decipher the molecular mechanisms that link NMD with translation termination.
The overall goal of this project is to comprehensively decipher the molecular events of premature translation termination that finally result in mRNA degradation by NMD. We will build on the experimental system that we have established or developed in the first funding period. This system includes a fully reconstituted in vitro translation system, selective ribosomal profiling, affinity purification of NMD factor-bound ribosomes followed by mass spectrometry (AP-MS), and structural studies. As specific aims, we will firstly elucidate the conditions of UPF3B function in translation termination in more depth and will directly probe the immediate function as well as the interaction network of SMG6 as a termination dependent mRNA endonuclease. Secondly, we will employ genome-edited cell lines to affinity-purify NMD factor-bound translating ribosomes followed (1) by mass spectrometry (AP-MS), (2) by ribosomal profiling, and (3) by cryo-electron microscopy thus contributing to a deeper understanding of tissue- and transcript-specific NMD branches and to the discovery of unknown protein:protein and protein:mRNA interactions at prematurely terminating ribosomes.
Project-related publications
Lavysh D, Neu-Yilik G.
UPF1-Mediated RNA Decay-Danse Macabre in a Cloud.
Biomolecules. 2020 Jul 4;10(7):999. doi: 10.3390/biom10070999
Neu-Yilik G, Raimondeau E, Eliseev B, Yeramala L, Amthor B, Deniaud A, Huard K, Kerschgens K, Hentze MW, Schaffitzel C, Kulozik AE.
Dual function of UPF3B in early and late translation termination. EMBO J. 2017 Oct 16;36(20):2968-2986. doi: 10.15252/embj.201797079
Deniaud A, Karuppasamy M, Bock T, Masiulis S, Huard K, Garzoni F, Kerschgens K, Hentze MW, Kulozik AE, Beck M, Neu-Yilik G, Schaffitzel C.
A network of SMG-8, SMG-9 and SMG-1 C-terminal insertion domain regulates UPF1 substrate recruitment and phosphorylation. Nucleic Acids Res. 2015 Sep 3;43(15):7600-11. doi: 10.1093/nar/gkv668