A5005089660- Ubiquitin and proteasome pathways
- Cellular Mechanics and Interactions
- Biochemical and Molecular Research
- Glycosylation and Glycoproteins Research
- RNA Research and Splicing
- Carbohydrate Chemistry and Synthesis
- Autophagy in Disease and Therapy
- Enzyme Structure and Function
- RNA modifications and cancer
- Endoplasmic Reticulum Stress and Disease
- Mitochondrial Function and Pathology
- Microtubule and mitosis dynamics
- Cancer, Hypoxia, and Metabolism
- Amino Acid Enzymes and Metabolism
- Cell Adhesion Molecules Research
- Genomics and Chromatin Dynamics
- Cellular transport and secretion
- Polyamine Metabolism and Applications
- Cancer Research and Treatments
- Signaling Pathways in Disease
- Hippo pathway signaling and YAP/TAZ
- Advanced Fluorescence Microscopy Techniques
- S100 Proteins and Annexins
- Cancer-related Molecular Pathways
- Protein Kinase Regulation and GTPase Signaling
The Francis Crick Institute
2016-2025
Cancer Research UK
2008-2014
The Honourable Society of Lincoln's Inn
2011-2012
Institut de Chimie des Substances Naturelles
2005-2012
Centre National de la Recherche Scientifique
2007-2012
Laboratoire d'Enzymologie et Biochimie Structurales
2005-2012
Birkbeck, University of London
2012
Institute of Structural and Molecular Biology
2012
Hokkaido University
2011
AgResearch
2008
The Atg8 family of ubiquitin-like proteins play pivotal roles in autophagy and other processes involving vesicle fusion transport where the lysosome/vacuole is end station. Nuclear are also emerging. Here, we review structural functional features their protein-protein interaction modes model organisms such as yeast, Arabidopsis, C. elegans Drosophila to humans. Although varying number homologs, from one yeast seven humans, more than ten some plants, there a strong evolutionary conservation...
Abstract Autophagy is an essential recycling and quality control pathway. Mammalian ATG8 proteins drive autophagosome formation selective removal of protein aggregates organelles by recruiting autophagy receptors adaptors that contain a LC3-interacting region (LIR) motif. LIR motifs can be highly for subfamily (LC3s/GABARAPs), however the molecular determinants regulating these interactions remain elusive. Here we show residues within core motif adjacent C-terminal as well subfamily-specific...
Autophagosome formation depends on a carefully orchestrated interplay between membrane-associated protein complexes. Initiation of macroautophagy/autophagy is mediated by the ULK1 (unc-51 like autophagy activating kinase 1) complex and autophagy-specific class III phosphatidylinositol 3-kinase I (PtdIns3K-C1). The latter contains PIK3C3/VPS34, PIK3R4/VPS15, BECN1/Beclin 1 ATG14 phosphorylates to generate 3-phosphate (PtdIns3P). Here, we show that PIK3C3, BECN1 contain functional LIR motifs...
The cysteine protease ATG4B cleaves off one or more C-terminal residues of the inactive proform proteins ortholog and paralog LC3 GABARAP subfamilies yeast Atg8 to expose a glycine that is conjugated phosphatidylethanolamine during autophagosome formation. We show contains LC3-interacting region (LIR) motif important for efficient binding cleavage proteins. solved crystal structures GABARAPL1-ATG4B LIR complex. Analyses in vitro assays, using specific point mutants, clearly showed binds via...
Mitophagy is the degradation of surplus or damaged mitochondria by autophagy. In addition to programmed and stress-induced mitophagy, basal mitophagy processes exert organelle quality control. Here, we show that sorting assembly machinery (SAM) complex protein SAMM50 interacts directly with ATG8 family proteins p62/SQSTM1 act as a receptor for components SAM mitochondrial contact site cristae organizing system (MICOS) complexes. regulates architecture controlling formation MICOS decisive...
Subcellular localization of the actin-binding transcriptional coactivator MRTF-A is controlled by its interaction with monomeric actin (G-actin). Signal-induced decreases in G-actin concentration reduce nuclear export, leading to accumulation, whereas artificial increases resting cells block import, retaining it cytoplasm. This regulation dependent on three RPEL motifs regulatory domain MRTF-A. We describe structures pentavalent and trivalent G-actin•RPEL complexes. In complex, each motif...
Abstract The restrictor, ZC3H4/WDR82, is the major termination factor for antisense transcription from bidirectional promoters, but its mechanism poorly understood. We report that ZC3H4/WDR82 co-purifies with PP1 phosphatase and nuclear targeting subunit, PNUTS, which binds directly to WDR82 subunit of restrictor. AlphaFold predicts a quaternary complex, PPWZ, in P P1-associated NUTS Z C3H4 both contact W DR82. To investigate role protein dephosphorylation PPWZ activity, we expressed...
Highlights•Kinetics of RET autophosphorylation identify early and late sites•Evidence for a kinase domain cis-inhibitory tether is presented•Oncogenic subverts cis-inhibition perturbs the trajectory•Oncogenic overactive better trans-autophosphorylation substrateSummaryTo decipher molecular basis activation oncogenic deregulation, we defined temporal sequence by label-free quantitative mass spectrometry. Early sites map to regions flanking core, while within loop only form at later time...
The Phactr family of PP1-binding proteins is implicated in human diseases including Parkinson's, cancer and myocardial infarction. Each protein contains four G-actin binding RPEL motifs, an N-terminal motif, abutting a basic element, C-terminal triple repeat, which overlaps conserved C-terminus required for interaction with PP1. motifs are also found the regulatory domains MRTF transcriptional coactivators, where they control subcellular localisation activity by sensing signal-induced...
Abstract Serine/threonine phosphatases such as PP1 lack substrate specificity and associate with a large array of targeting subunits to achieve the requisite selectivity. The tumour suppressor ASPP (apoptosis-stimulating protein p53) proteins catalytic are implicated in multiple functions from transcriptional regulation cell junction remodelling. Here we show that Drosophila is part multiprotein complex association necessary for several vivo ASPP. We solve crystal structure human ASPP2/PP1...
The restrictor, ZC3H4/WDR82, terminates antisense transcription from bidirectional promoters, but its mechanism is poorly understood. We report that ZC3H4/WDR82 immunoprecipitate with PP1 phosphatase and nuclear targeting subunit, PNUTS, which binds to WDR82. AlphaFold predicts a complex of PP1/PNUTS restrictor where both PNUTS ZC3H4 contact A substrate trap, PP1H66K-PNUTS, comprising inactive fused the C-terminus antagonizes mediated termination whereas PP1WT-PNUTS has less effect...
Glucosamine-6P synthase catalyzes the synthesis of glucosamine-6P from fructose-6P and glutamine uses a channel to transfer ammonia its glutaminase active site. X-ray structures have been determined at 2.05 Angstroms resolution in presence 2.35 6-diazo-5-oxo-L-norleucine, affinity analog that covalently modifies N-terminal catalytic cysteine, therefore mimicking gamma-glutamyl-thioester intermediate formed during hydrolysis glutamine. The fixation activates enzyme through several major...
Receptor tyrosine kinases exhibit a variety of activation mechanisms despite highly homologous catalytic domains. Such diversity arises through coupling extracellular ligand-binding portions with variable intracellular sequences flanking the kinase domain and specific patterns autophosphorylation sites. Here, we show that juxtamembrane (JM) segment enhances RET activity Y687. This phospho-site is also required by JM region to rescue an otherwise catalytically deficient activation-loop mutant...
PPP-family phosphatases such as PP1 have little intrinsic specificity. Cofactors can target to substrates or subcellular locations, but it remains unclear how they might confer sequence-specificity on PP1. The cytoskeletal regulator Phactr1 is a neuronally enriched cofactor that controlled by G-actin. Structural analysis showed binding remodels PP1's hydrophobic groove, creating new composite surface adjacent the catalytic site. Using phosphoproteomics, we identified mouse fibroblast and...
Autophagy is a highly conserved degradative pathway, essential for cellular homeostasis and implicated in diseases including cancer neurodegeneration. Autophagy-related 8 (ATG8) proteins play central role autophagosome formation selective delivery of cytoplasmic cargo to lysosomes by recruiting autophagy adaptors receptors. The LC3-interacting region (LIR) docking site (LDS) ATG8 binds LIR motifs present LIR-ATG8 interactions can be specific mammalian family members (LC3A-C, GABARAP,...
The structure-specific endonuclease activity of the human XPF–ERCC1 complex is essential for a number DNA processing mechanisms that help to maintain genomic integrity. cleaves structures such as stem–loops, bubbles or flaps in one strand duplex where there at least downstream single strand. Here, we define minimal substrate requirements cleavage stem–loop substrates allowing us develop real-time fluorescence-based assay measure activity. Using this assay, show changes sequence upstream...
Ammonia transfer from the glutamine site to fructose-6P of bacterial glucosamine-6-phosphate synthase was studied by molecular dynamics simulations. The studies suggest a key role for Trp74, in sealing hydrophobic channel connecting two binding sites, as well Ala602 and Val605 residues, which form narrow passage whose opening/closing constitutes an essential event ammonia transfer. Kinetic analyses corresponding protein mutants confirmed our predictions. efficiency close zero W74A mutant...