An international team of researchers from the Max Planck Research Group at the Malopolska Centre of Biotechnology (MCB) deciphered the molecular mechanism of a sulfur carrier system that stands at the root of all eukaryotic Ubiquitin-conjugation systems. For this project, the team headed by Sebastian Glatt from the Jagiellonian University in Krakow (Poland) collaborated closely with the group of Sebastian Leidel at the Department of Chemistry and Biochemistry of the University of Bern (Switzerland). The results of their research were recently published in “The EMBO Journal”.
The study provides the first structural and mechanistic insight into the eukaryotic Uba4-Urm1 system, which is essential for the transfer and incorporation of sulfur into transfer RNA (tRNA) bases. Perturbation of these specific modification pathways results in protein homeostasis defects and has been linked to neurological disorders and cancer in humans.
The team obtained the first three-dimensional atomic model of the E1-like sulfurtransferase, Uba4, with its bound substrate, the Ubiquitin-like protein (UBL), Urm1. The crystal structure of the Uba4-Urm1 complex provides molecular insights into substrate recognition and the enzymatic reaction cascade. In addition, the authors decipher a critical role of an active site residue that protects Uba4 from its own reaction product. As this feature appears to be conserved in all other E1 and E1-like activating domains, the observed covalent E1-UBL self-conjugation reaction likely represents the evolutionary ancestor of most UBL pathways. The discovery of the Ubiquitin-mediated proteolysis has received the Nobel Prize in 2004 and involves thousands of cellular proteins. UBLs and the underlying degradation pathways have been the focus of many academic and industrial projects, aiming to exploit them as diagnostic markers, drug targets and molecular drugs themselves.
The lead author, Marta Pabis, explains that “UBL protein modifications in eukaryotes might have evolved from prokaryotic sulfurtransferase systems and our study provides further evidence that the Uba4-Urm1 system represents that missing ‘molecular fossil’“. The second lead author from the MCB, Keerthiraju E. Ravichandran, adds “with great power comes great responsibility – the Uba4-Urm1 system has taken its responsibility and protected itself from its own product. Thereby, it allowed the creation of a whole universe of novel cellular signalling pathways.” Sebastian Glatt summarizes “our work provides a milestone in understanding the dual character of this system and provides deep molecular insights in the underlying mechanisms.”
The work was mainly supported by the First Team Grant (FirstTEAM/2016-1/2) from the Foundation for Polish Science. In addition, the MCB Structural Biology Core Facility (supported by the TEAM TECH CORE FACILITY/2017-4/6 grant from Foundation for Polish Science) provided crucial support and essential equipment.
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