A5067524265- Ubiquitin and proteasome pathways
- DNA Repair Mechanisms
- Genetics and Neurodevelopmental Disorders
- 14-3-3 protein interactions
- Autophagy in Disease and Therapy
- Microtubule and mitosis dynamics
- CRISPR and Genetic Engineering
- Mitochondrial Function and Pathology
- Protein Kinase Regulation and GTPase Signaling
- Cancer-related Molecular Pathways
- Cellular transport and secretion
- Plant Virus Research Studies
- Genomics and Chromatin Dynamics
- Protein Tyrosine Phosphatases
- Parkinson's Disease Mechanisms and Treatments
- RNA Research and Splicing
- Protein Degradation and Inhibitors
- Pluripotent Stem Cells Research
- PARP inhibition in cancer therapy
- Peptidase Inhibition and Analysis
- interferon and immune responses
- Transgenic Plants and Applications
- Cytokine Signaling Pathways and Interactions
- Microbial Natural Products and Biosynthesis
- Cell death mechanisms and regulation
MRC Protein Phosphorylation and Ubiquitylation Unit
2016-2025
University of Dundee
2016-2025
Wellcome Trust
2018-2024
Medical Research Council
2008-2022
Cleveland Clinic
2017-2019
Czech Agrifood Research Center
2003
University of Warwick
1999
In this study, we develop a simple assay to identify mitophagy inducers on the basis of use fluorescently tagged mitochondria that undergo colour change lysosomal delivery. Using assay, iron chelators as family compounds generate strong response. Iron chelation‐induced requires cells glycolysis, but does not require PINK1 stabilization or Parkin activation, and occurs in primary human fibroblasts well those isolated from Parkinson's patient with mutations. Thus, have identified characterized...
More than 200 phosphorylated 14-3-3-binding sites in the literature were analysed to define 14-3-3 specificities, identify relevant protein kinases, and give insights into how cellular 14-3-3/phosphoprotein networks work. Mode I RXX(pS/pT)XP motifs dominate, although +2 proline residue occurs less half, LX(R/K)SX(pS/pT)XP is prominent plant sites. Proline at +1 rarely reported, such did not stand up experimental reanalysis of human Ndel1. Instead, we discovered that interacts with two...
The polarization of macrophages into a regulatory-like phenotype and the production IL-10 plays an important role in resolution inflammation. We show this study that PGE2, combination with LPS, is able to promote anti-inflammatory characterized by high expression regulatory markers SPHK1 LIGHT via protein kinase A–dependent pathway. Both TLR agonists PGE2 phosphorylation transcription factor CREB on Ser133. However, although regulates transcription, mutation Ser133 Ala endogenous gene did...
Abstract Motivation: The 14-3-3 family of phosphoprotein-binding proteins regulates many cellular processes by docking onto pairs phosphorylated Ser and Thr residues in a constellation intracellular targets. Therefore, there is pressing need to develop new prediction methods that use an updated set 14-3-3-binding motifs for the identification targets prioritize downstream analysis >2000 potential interactors identified high-throughput experiments. Results: Here, comprehensive from...
Abstract The PARK 2 gene is mutated in 50% of autosomal recessive juvenile parkinsonism ( ARJP ) cases. It encodes parkin, an E3 ubiquitin ligase the RBR family. Parkin exists autoinhibited state that activated by phosphorylation its N‐terminal ubiquitin‐like (Ubl) domain and binding phosphoubiquitin. We describe 1.8 Å crystal structure human parkin fully inhibited identify key interfaces to maintain inhibition. phosphoubiquitin‐binding interface, provide a model for phosphoubiquitin–parkin...
Abstract Background Pim-1, 2 and 3 are a group of enzymes related to the calcium calmodulin family protein kinases. Over-expression Pim-1 Pim-2 in mice promotes development lymphomas, up-regulation Pim expression has been observed several human cancers. Results Here we show that pim kinases constitutively active when expressed HEK-293 cells able phosphorylate Bcl-2 member Bad on three residues, Ser112, Ser136 Ser155 vitro cells. In mapping showed predominantly phosphorylated while but also...
AS160 (Akt substrate of 160 kDa) and TBC1D1 are related RabGAPs (Rab GTPase-activating proteins) implicated in regulating the trafficking GLUT4 (glucose transporter 4) storage vesicles to cell surface. All animal species examined contain TBC1D1, whereas evolved with vertebrates. has two clusters phosphorylated residues, either side second PTB (phosphotyrosine-binding domain). Each cluster contains a 14-3-3-binding site. When AMPK (AMP-activated protein kinase) is activated HEK (human...
AS160 (Akt substrate of 160 kDa) mediates insulin-stimulated GLUT4 (glucose transporter 4) translocation, but is widely expressed in insulin-insensitive tissues lacking GLUT4. Having isolated by 14-3-3-affinity chromatography, we found that binding to 14-3-3 isoforms HEK (human embryonic kidney)-293 cells was induced IGF-1 (insulin-like growth factor-1), EGF (epidermal factor), PMA and, a lesser extent, AICAR (5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside). AS160-14-3-3 interactions...
Article2 July 2015Open Access Source Data mTOR activates the VPS34–UVRAG complex to regulate autolysosomal tubulation and cell survival Michael J Munson MRC Protein Phosphorylation Ubiquitylation Unit, College of Life Sciences, University Dundee, UK Search for more papers by this author George FG Allen Rachel Toth David G Campbell John M Lucocq School Medicine, St Andrews, Ian Ganley Corresponding Author Information Munson1, Allen1, Toth1, Campbell1, Lucocq2 1 1MRC 2School *Corresponding...
The protein ABIN1 possesses a polyubiquitin-binding domain homologous to that present in nuclear factor κB (NF-κB) essential modulator (NEMO), component of the inhibitor NF-κB (IκB) kinase (IKK) complex. To address physiological significance polyubiquitin binding, we generated knockin mice expressing ABIN1[D485N] mutant instead wild-type (WT) protein. These developed all hallmarks autoimmunity, including spontaneous formation germinal centers, isotype switching, and production autoreactive...
Polyubiquitin chains regulate diverse cellular processes through the ability of ubiquitin to form eight different linkage types. Although detected in yeast and mammals, little is known about K29-linked polyubiquitin. Here we report generation K29 vitro using a chain-editing complex consisting HECT E3 ligase UBE3C deubiquitinase vOTU. We determined crystal structure diubiquitin, which adopts an extended conformation with hydrophobic patches on both moieties exposed available for binding....
Holliday junctions (HJs) are X-shaped DNA structures that arise during homologous recombination, which must be removed to enable chromosome segregation. The SLX1 and MUS81-EME1 nucleases can both process HJs in vitro, they bind close proximity on the SLX4 scaffold, hinting at possible cooperation. However, cellular roles of mammalian not yet known. Here, we use mouse genetics structure function analysis investigate function. Disrupting murine Slx1 Slx4 genes revealed essential for HJ...
Mono-ubiquitination of Fancd2 is essential for repairing DNA interstrand cross-links (ICLs), but the underlying mechanisms are unclear. The Fan1 nuclease, also required ICL repair, recruited to ICLs by ubiquitinated (Ub) Fancd2. This could in principle explain how Ub-Fancd2 promotes we show that recruitment dispensable repair. Instead, recruitment--and activity--restrains replication fork progression and prevents chromosome abnormalities from occurring when forks stall, even absence ICLs....
Of the 16 non-structural proteins (Nsps) encoded by SARS CoV-2, Nsp3 is largest and plays important roles in viral life cycle. Being a large, multidomain, transmembrane protein, has been most challenging Nsp to characterize. Encoded within papain-like protease domain (PLpro) that cleaves not only polypeptide but also K48-linked polyubiquitin ubiquitin-like modifier, ISG15, from host cell proteins. We here compare interactors of PLpro find largely overlapping interactome. Intriguingly, we...
Loss-of-function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinson’s disease (PD). Stabilization PINK1 at the translocase outer membrane (TOM) complex damaged mitochondria is critical for its activation. The mechanism how activated TOM unclear. Here, we report that co-expression human and all seven subunits Saccharomyces cerevisiae sufficient We use this reconstitution system to systematically assess role each subunit toward unambiguously demonstrate...
Microtubules interact strongly with the viral movement protein (MP) of Tobacco mosaic virus (TMV) and are thought to transport genome between plant cells. We describe a functionally enhanced DNA-shuffled (MP(R3)) that remained bound vertices cortical endoplasmic reticulum, showing limited affinity for microtubules. A single amino acid change was shown confer MP(R3) phenotype. Disruption microtubule cytoskeleton in situ pharmacological agents, or by silencing alpha-tubulin gene, had no...
Recent work indicates that the LKB1 tumour suppressor protein kinase, which is mutated in Peutz–Jeghers cancer syndrome, phosphorylates and activates a group of kinases are related to AMPK (AMP-activated kinase). Ten 14 AMPK-related activated by LKB1, including SIK (salt-induced kinase), MARK (microtubule-affinity-regulating kinase) BRSK (brain-specific isoforms, possess ubiquitin-associated (UBA) domain immediately C-terminal kinase catalytic domain. These only human genome known UBA...
Compounds that inhibit signalling upstream of ERK (extracellular-signal-regulated kinase) are promising anticancer therapies, motivating research to define how this pathway promotes cancers. In the present study, we show human capicúa represses mRNA expression for PEA3 (polyoma enhancer activator 3) Ets transcription factors ETV1, ETV4 and ETV5 (ETV is translocation variant), repression relieved by multisite controls ERK, p90(RSK) (p90 ribosomal S6 14-3-3 proteins. Specifically, binds...
ATM (ataxia-telangiectasia mutated), ATR (ATM- and Rad3-related) DNA-PK (DNA-dependent protein kinase), important regulators of genome stability, belong to the PIKK (phosphoinositide 3-kinase-like kinase) family kinases. In present study, DNA-affinity chromatography was used identify DNA-binding proteins phosphorylated by these This resulted in identification FUS (fused sarcoma)/TLS (translocated liposarcoma) as an vitro target PIKKs. is a member Ewing's sarcoma that appears play role...
Human (h)PTIP plays important but poorly understood roles in cellular responses to DNA damage. hPTIP interacts with 53BP1 tumour suppressor only when is phosphorylated by ATM after damage although the mechanism(s) and significance of interaction these two proteins are unclear. Here, we pinpoint a single ATM-phosphorylated residue 53BP1--Ser25--that required for binding hPTIP. Binding phospho-Ser25 vitro vivo requires closely apposed pairs BRCT domains at C-terminus neither pair alone can...
SLX4, a scaffold for structure-specific DNA repair nucleases, is important several types of repair. Many proteins bind to sites damage, resulting in subnuclear "foci," but SLX4 forms foci human cells even without damage. Using approaches, we show that most, not all, localize telomeres range cell lines irrespective the mechanisms used maintain telomere length. The SLX1 Holliday-junction-processing enzyme recruited by and turn, motif binds shelterin subunit TRF2 directly. We also...