A5069109050- Ubiquitin and proteasome pathways
- Protein Degradation and Inhibitors
- RNA and protein synthesis mechanisms
- Biochemical and Molecular Research
- Glycosylation and Glycoproteins Research
- RNA modifications and cancer
- 14-3-3 protein interactions
- Tuberculosis Research and Epidemiology
- Cancer-related Molecular Pathways
- Endoplasmic Reticulum Stress and Disease
- Enzyme Structure and Function
- Mycobacterium research and diagnosis
- Autophagy in Disease and Therapy
- Microtubule and mitosis dynamics
- Cancer, Hypoxia, and Metabolism
- Cellular transport and secretion
- Photosynthetic Processes and Mechanisms
- RNA regulation and disease
- Peptidase Inhibition and Analysis
- RNA Research and Splicing
- DNA Repair Mechanisms
- Genetics, Bioinformatics, and Biomedical Research
- Advanced Proteomics Techniques and Applications
- interferon and immune responses
- Cancer therapeutics and mechanisms
Max Planck Institute of Biochemistry
2014-2024
Max Planck Society
2015-2020
ETH Zurich
2020
Deutsche Forschungsgemeinschaft
2020
Indian Institute of Science Bangalore
2006-2015
Abstract E3 ligases are typically classified by hallmark domains such as RING and RBR, which thought to specify unique catalytic mechanisms of ubiquitin transfer recruited substrates 1,2 . However, rather than functioning individually, many neddylated cullin–RING (CRLs) RBR-type in the ARIH family—which together account for nearly half all humans—form E3–E3 super-assemblies 3–7 Here, studying CRLs SKP1–CUL1–F-box (SCF) family, we show how SCF ARIH1 (an ligase) co-evolved ubiquitylate diverse...
Highlights•UBE2R E2s preferentially target CUL2-based CRL2 substrates•CRL2-dependent PROTAC efficiency correlates with UBE2R-family E2 levels in cells•Cryo-EM reveals a UBE2R2-CRL2 interface that underlies geometric substrate selection•A dedicated poly-ubiquitin chain extending is reprogrammed to prime substratesSummaryCullin-RING ligases (CRLs) ubiquitylate specific substrates selected from other cellular proteins. Substrate discrimination and ubiquitin transferase activity were thought be...
Membrane protein biogenesis in the endoplasmic reticulum (ER) is complex and failure-prone. The ER membrane (EMC), comprising eight conserved subunits, has emerged as a central player this process. Yet, we have limited understanding of how EMC enables insertion integrity diverse clients, from tail-anchored to polytopic transmembrane proteins. Here, yeast human cryo-EM structures reveal intricate assemblies human-specific features associated with pathologies. Structure-based functional...
Ubiquitin (Ub) chain formation by homologous to E6AP C-terminus (HECT)-family E3 ligases regulates vast biology, yet the structural mechanisms remain unknown. We used chemistry and cryo-electron microscopy (cryo-EM) visualize stable mimics of intermediates along K48-linked Ub human E3, UBR5. The data reveal a ≈ 620 kDa UBR5 dimer as functional unit, comprising scaffold with flexibly tethered Ub-associated (UBA) domains, elaborately arranged HECT domains. Chains are forged UBA domain...
Abstract E3 ubiquitin ligases, in collaboration with E2 ubiquitin-conjugating enzymes, modify proteins poly-ubiquitin chains. Cullin-RING ligase (CRL) E3s use Cdc34/UBE2R-family E2s to build Lys48-linked chains control an enormous swath of eukaryotic biology. Yet the molecular mechanisms underlying this exceptional linkage specificity and millisecond kinetics poly-ubiquitylation remain unclear. Here we obtain cryogenic-electron microscopy (cryo-EM) structures that provide pertinent insight...
Abstract An emerging mechanism of ubiquitylation involves partnering two distinct E3 ligases. In the best-characterized E3-E3 pathways, ARIH-family RING-between-RING (RBR) E3s ligate ubiquitin to substrates neddylated cullin-RING E3s. The ARIH2 has been implicated in CUL5-RBX2-based E3s, including APOBEC3-family host hijacked by HIV-1 virion infectivity factor (Vif). However, structural mechanisms remained elusive. Here and biochemical analyses reveal distinctive autoinhibition, activation...
The yeast THO complex is recruited to active genes and interacts with the RNA-dependent ATPase Sub2 facilitate formation of mature export-competent messenger ribonucleoprotein particles prevent co-transcriptional RNA:DNA-hybrid-containing structures. How THO-containing complexes function at mechanistic level unclear. Here, we elucidated a 3.4 Å resolution structure Saccharomyces cerevisiae THO-Sub2 by cryo-electron microscopy. subunits Tho2 Hpr1 intertwine form platform that bound Mft1,...
N-degron E3 ubiquitin ligases recognize specific residues at the N-termini of substrates. Although molecular details recognition are known for several ligases, range N-terminal motifs that can bind a given substrate binding domain remains unclear. Here, we discovered capacity Gid4 and Gid10 receptor subunits yeast "GID"/human "CTLH" multiprotein to tightly wide whose is determined in part by downstream sequence context. Screening phage displaying peptide libraries with exposed identified...
Ubiquitylation is catalyzed by coordinated actions of E3 and E2 enzymes. Molecular principles governing many important E3-E2 partnerships remain unknown, including those for RING-family GID/CTLH ubiquitin ligases their dedicated E2, Ubc8/UBE2H (yeast/human nomenclature). GID/CTLH-Ubc8/UBE2H-mediated ubiquitylation regulates biological processes ranging from yeast metabolic signaling to human development. Here, cryoelectron microscopy (cryo-EM), biochemistry, cell biology reveal this...
Ubiquitin (Ub)-mediated proteolysis is a fundamental mechanism used by eukaryotic cells to maintain homeostasis and protein quality, control timing in biological processes. Two essential aspects of Ub regulation are conjugation through E1-E2-E3 enzymatic cascades recognition Ub-binding domains. An emerging theme the field that these 2 properties often amalgamated enzymes. In addition covalent thioester linkage Ub’s C terminus for transfer reactions, enzymes bind noncovalently weakly at...
Abstract Ubiquitin ligation is typically executed by hallmark E3 catalytic domains. Two such domains, ‘cullin–RING’ and ‘RBR’, are individually found in several hundred human ligases, collaborate with E2 enzymes to catalyze ubiquitylation. However, the vertebrate-specific CUL9 complex RBX1 (also called ROC1), of interest due its tumor suppressive interaction TP53, uniquely encompasses both cullin–RING RBR Here, cryo-EM, biochemistry cellular assays elucidate a 1.8-MDa hexameric CUL9–RBX1...
Abstract The attachment of the ubiquitin-like protein ISG15 to substrates by specific E1-E2-E3 enzymes is a well-established signalling mechanism innate immune response. Here, we present 3.45 Å cryo-EM structure chemically trapped UBE1L-UBE2L6 complex bound activated ISG15. This reveals details first steps recognition and UBE2L6 recruitment UBE1L (also known as UBA7). Taking advantage viral effector proteins from severe acute respiratory coronavirus 2 (SARS-CoV-2) influenza B virus (IBV),...
Abstract The attachment of the ubiquitin-like protein ISG15 to substrates is a well-established antiviral signalling mechanism innate immune response. However, despite identification thousands and clear roles in immunity, molecular understanding selection transfer through its cognate E1-E2- E3 enzyme cascade largely unknown. Here, we present 3.45 Å cryo-EM structure chemically trapped UBE1L-UBE2L6 complex bound activated ISG15. This reveals details first steps recognition UBE2L6 recruitment...
ABSTRACT Ubiquitin chain formation by HECT catalytic domain-containing E3 ligases regulates vast biology, yet the structural mechanisms remain unknown. We employed chemistry and cryo-EM to visualize stable mimics of intermediates along K48-linked ubiquitin human E3, UBR5. The data reveal a ≈620 kDa UBR5 dimer as functional unit, comprising scaffold with flexibly-tethered ubiquitin-binding UBA domains, elaborately arranged domains. Chains are forged domain capturing an acceptor ubiquitin, its...