A5018035568- Virology and Viral Diseases
- Cytomegalovirus and herpesvirus research
- HIV Research and Treatment
- Bacteriophages and microbial interactions
- Mosquito-borne diseases and control
- Viral Infections and Vectors
- CRISPR and Genetic Engineering
- Hepatitis B Virus Studies
- Viral gastroenteritis research and epidemiology
- HIV/AIDS drug development and treatment
- Microtubule and mitosis dynamics
- Virus-based gene therapy research
- Advanced Electron Microscopy Techniques and Applications
- Glycogen Storage Diseases and Myoclonus
- Lipid Membrane Structure and Behavior
- Photosynthetic Processes and Mechanisms
- Herpesvirus Infections and Treatments
- RNA modifications and cancer
- Biochemical and Molecular Research
- Carbohydrate Chemistry and Synthesis
- Nanopore and Nanochannel Transport Studies
- Mitochondrial Function and Pathology
- Viral-associated cancers and disorders
- Plant Virus Research Studies
- Protein purification and stability
Institute of Structural and Molecular Biology
2022-2024
University of Leeds
2017-2024
The Francis Crick Institute
2015-2019
Cancer Research UK
2010
Retroviral integrase (IN) functions within the intasome nucleoprotein complex to catalyze insertion of viral DNA into cellular chromatin. Using cryo-electron microscopy, we now visualize functional maedi-visna lentivirus at 4.9 angstrom resolution. The comprises a homo-hexadecamer IN with tetramer-of-tetramers architecture featuring eight structurally distinct types protomers supporting two catalytically competent subunits. conserved intasomal core, previously observed in simpler retroviral...
The MIND multiprotein complex is a conserved, essential component of eukaryotic kinetochores and constituent the tripartite KMN network that directly attaches kinetochore to mitotic spindle. primary microtubule-binding in this network, NDC80, has been extensively characterized, but very little known about structure or function complex. In study, we present biochemical, hydrodynamic, electron microscopy, small-angle x-ray scattering data provide insight into overall architecture assembly...
Abstract Insect pests are a major cause of crop losses worldwide, with an estimated economic cost $470 billion annually. Biotechnological tools have been introduced to control such insects without the need for chemical pesticides; instance, development transgenic plants harbouring genes encoding insecticidal proteins. The Vip3 (vegetative protein 3) family proteins from Bacillus thuringiensis convey toxicity species within Lepidoptera, and wide potential applications in commercial...
An artificial protein cage can be programmed to disassemble on demand.
Abstract Glycogen is the major glucose reserve in eukaryotes, and defects glycogen metabolism structure lead to disease. Glycogenesis involves interaction of glycogenin (GN) with synthase (GS), where GS activated by glucose-6-phosphate (G6P) inactivated phosphorylation. We describe 2.6 Å resolution cryo-EM phosphorylated human revealing an autoinhibited tetramer flanked two GN dimers. Phosphorylated N- C-termini from protomers converge near G6P-binding pocket buttress against regulatory...
The interactions between a retrovirus and host cell chromatin that underlie integration provirus expression are poorly understood. prototype foamy virus (PFV) structural protein GAG associates with chromosomes via chromatin-binding sequence (CBS) located within its C-terminal region. Here, we show the PFV CBS is essential sufficient for direct interaction nucleosomes present crystal structure of bound to mononucleosome. interacts histone octamer, engaging H2A-H2B acidic patch in manner...
Integrase mutations can reduce the effectiveness of first-generation FDA-approved integrase strand transfer inhibitors (INSTIs), raltegravir (RAL) and elvitegravir (EVG). The second-generation agent, dolutegravir (DTG), has enjoyed considerable clinical success; however, resistance-causing that diminish efficacy DTG have appeared. Our current findings support extend substrate envelope concept broadly effective INSTIs be designed by filling defined DNA substrates. Previously, we explored...
Abstract Retroviral integrase can efficiently utilise nucleosomes for insertion of the reverse-transcribed viral DNA. In face structural constraints imposed by nucleosomal structure, gains access to scissile phosphodiester bonds lifting DNA off histone octamer at site integration. To clarify mechanism looping integrase, we determined a 3.9 Å resolution structure prototype foamy virus intasome engaged with nucleosome core particle. The data along complementary single-molecule Förster...
Flaviviruses , including Zika virus (ZIKV), are a significant global health concern, yet no licensed antivirals exist to treat disease. The small membrane (M) protein plays well-defined roles during viral egress and remains within virion membranes following release maturation. However, it is unclear whether M functional role in this setting. Here, we show that forms oligomeric membrane-permeabilising channels vitro, with increased activity at acidic pH sensitivity the prototypic...
HIV integrase (IN) strand transfer inhibitors (INSTIs) are among the newest anti-AIDS drugs; however, mutant forms of IN can confer resistance. We developed noncytotoxic naphthyridine-containing INSTIs that retain low nanomolar IC50 values against HIV-1 variants harboring all major INSTI-resistant mutations. found by analyzing crystal structures bound to from prototype foamy virus (PFV) most successful show striking mimicry viral DNA prior 3'-processing and host transfer. Using this concept...
Abstract Hybrid vesicles consisting of phospholipids and block‐copolymers are increasingly finding applications in science technology. Herein, small angle X‐ray scattering (SAXS) cryo‐electron tomography (cryo‐ET) used to obtain detailed structural information about hybrid with different ratios 1‐palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphocholine (POPC) poly(1,2‐butadiene‐block‐ethylene oxide) (PBd 22 ‐PEO 14 , M s = 1800 g mol −1 ). Using single particle analysis (SPA) the authors able further...
Abstract p27KIP1 (cyclin-dependent kinase inhibitor 1B, p27) is a member of the CIP/KIP family CDK (cyclin dependent kinase) regulators that inhibit cell cycle CDKs. p27 phosphorylation by CDK1/2, signals its recruitment to SCF SKP2 (S-phase associated protein 1 (SKP1)-cullin-SKP2) E3 ubiquitin ligase complex for proteasomal degradation. The nature binding and CKS1 was revealed SKP1-SKP2-CKS1-p27 phosphopeptide crystal structure. Subsequently, model hexameric CDK2-cyclin A-CKS1-p27-SKP1-SKP2...
Abstract We report the first cryoEM structure of Hendra henipavirus nucleoprotein in complex with RNA, at 3.5 Å resolution, derived from single particle analysis a double homotetradecameric RNA-bound N protein ring assembly exhibiting D14 symmetry. The HeV adopts common bi-lobed paramyxoviral fold; N-terminal and C-terminal globular domains are bisected by an RNA binding cleft containing six nucleotides flanked arms, respectively. In other nucleocapsids, lateral interface between adjacent i...
Abstract The roles of RNA sequence/structure motifs, Packaging Signals (PSs), for regulating assembly an HBV genome transcript have been investigated in efficient vitro assay containing only core protein (Cp) and RNA. Variants three conserved PSs, within the a strain not used previously, preventing correct presentation Cp-recognition loop motif are differentially deleterious nucleocapsid-like particles (NCPs). Cryo-electron microscopy reconstruction T = 4 NCPs formed with wild-type gRNA...
Flaviviruses , including Zika virus (ZIKV), are a significant global health concern, yet no licensed antivirals exist to treat disease. The small Membrane (M) protein plays well-defined roles during viral egress and remains within virion membranes following release maturation. However, it is unclear whether M functional role in this setting. Here, we show that forms oligomeric membrane-permeabilising channels vitro with increased activity at acidic pH sensitivity the prototypic...
Cryo-electron microscopy (cryoEM) is a powerful technique for structure determination of macromolecular complexes, via single particle analysis (SPA). The overall process involves i) vitrifying the specimen in thin film supported on cryoEM grid; ii) screening to assess distribution and ice quality; iii) if grid suitable, collecting dataset analysis; iv) image processing yield an EM density map. In this protocol, overview each these steps provided, with focus variables which user can modify...
Cryo-electron microscopy (cryoEM) is a powerful technique for structure determination of macromolecular complexes, via single particle analysis (SPA). The overall process involves i) vitrifying the specimen in thin film supported on cryoEM grid; ii) screening to assess distribution and ice quality; iii) if grid suitable, collecting dataset analysis; iv) image processing yield an EM density map. In this protocol, overview each these steps provided, with focus variables which user can modify...
Abstract We report the first cryoEM structure of Hendra henipavirus nucleoprotein in complex with RNA, at 3.5 Å resolution, derived from single particle analysis homotetradecameric RNA-bound N protein rings exhibiting D14 symmetry. The HeV adopts common bi-lobed paramyxoviral fold; N-terminal and C-terminal globular domains are bisected by an RNA binding cleft containing six nucleotides flanked arms, respectively. In other nucleocapsids, lateral interface between adjacent +1 protomers...