A5045735887- Bacteriophages and microbial interactions
- SARS-CoV-2 and COVID-19 Research
- Advanced Electron Microscopy Techniques and Applications
- Respiratory viral infections research
- Viral gastroenteritis research and epidemiology
- Viral Infections and Vectors
- Animal Virus Infections Studies
- Electron and X-Ray Spectroscopy Techniques
- Virology and Viral Diseases
- Bacterial biofilms and quorum sensing
- Receptor Mechanisms and Signaling
- Hepatitis B Virus Studies
- HIV Research and Treatment
- Virus-based gene therapy research
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Viral Infections and Immunology Research
- Plant Virus Research Studies
- Lipid Membrane Structure and Behavior
- Viral Infections and Outbreaks Research
- Advanced X-ray Imaging Techniques
- Cell Image Analysis Techniques
- Biochemical and Structural Characterization
- Advanced Fluorescence Microscopy Techniques
- CRISPR and Genetic Engineering
MRC Laboratory of Molecular Biology
2019-2025
Medical Research Council
2020-2025
The University of Texas at Austin
2024-2025
Max Planck Institute of Biochemistry
2021-2024
University Hospital Heidelberg
2021
Heidelberg University
2021
Children's Healthcare of Atlanta
2015-2020
Emory University
2015-2020
Georgia Institute of Technology
2013-2020
Wuhan University
2019
Elucidating a bacterial sense of touch Bacteria can adhere to surfaces within the host. This leads tissue colonization, induction virulence, and eventually formation biofilms—multicellular communities that resist antibiotics clearance by immune system (see Perspective Hughes Berg). Hug et al. show bacteria have allows them change their behavior rapidly when encountering surfaces. tactile sensing makes use inner components flagellum, rotary motor powered proton motif force facilitates...
We present a new approach for macromolecular structure determination from multiple particles in electron cryo-tomography (cryo-ET) data sets. Whereas existing subtomogram averaging approaches are based on 3D models, we propose to optimise regularised likelihood target that approximates function of the 2D experimental images. In addition, analogous Bayesian polishing and contrast transfer (CTF) refinement single-particle analysis, describe exploit increased signal-to-noise ratio averaged...
Abstract Measles virus (MeV) remains a major human pathogen, but there are presently no licensed antivirals to treat MeV or other paramyxoviruses. Here, we use cryo-electron tomography (cryo-ET) elucidate the principles governing paramyxovirus assembly in MeV-infected cells. The three-dimensional (3D) arrangement of structural proteins including surface glycoproteins (F and H), matrix protein (M), ribonucleoprotein complex (RNP) characterized at stages budding, released particles. M is...
Abstract Respiratory syncytial virus (RSV) is a leading cause of infant hospitalization and there remains no pediatric vaccine. RSV live-attenuated vaccines (LAVs) have history safe testing in infants; however, achieving an effective balance attenuation immunogenicity has proven challenging. Here we seek to engineer LAV with enhanced immunogenicity. Genetic mapping identifies strain line 19 fusion (F) protein residues that correlate pre-fusion antigen maintenance by ELISA thermal stability...
Structural changes in HIV maturation Nascent particles assemble at the plasma membrane of an infected cell and bud into a membrane-enveloped, immature virion. Assembly budding are driven by polyprotein called Gag, which consists matrix domain (MA) that is recruited to membrane, capsid (CA) responsible for self-assembly, nucleocapsid (NC) recruits viral RNA genome. Gag cleavage results structural rearrangement produces mature Qu et al . imaged electron tomography focused on MA (see...
Human respiratory syncytial virus (RSV) is the leading cause of lower tract disease in young children. With repeat infections throughout life, it can also substantial elderly and adults with compromised cardiac, pulmonary immune systems. RSV a pleomorphic enveloped RNA Pneumoviridae family. Recently, three-dimensional (3D) structure purified particles has been elucidated, revealing three distinct morphological categories: spherical, asymmetric, filamentous. However, native 3D associated or...
HIV immature lattice stability affects capsid maturation by altering IP 6 packaging.
The molecular events that permit the spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to bind and enter cells are important understand for both fundamental therapeutic reasons. Spike proteins consist S1 S2 domains, which recognize angiotensin-converting enzyme (ACE2) receptors contain viral fusion machinery, respectively. Ostensibly, binding trimers ACE2 promotes dissociation domains exposure although details this process have yet be observed. We report...
Reconstitution of the PMP22 protein into lipid bilayers results in membrane assemblies that share common features with myelin.
Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions are surrounded by a lipid bilayer from which spike (S) protein trimers protrude. Heavily glycosylated S bind the ACE2 receptor and mediate entry of into target cells. exhibits extensive conformational flexibility: it modulates exposure its binding site later undergoes complete structural rearrangement to drive fusion viral cellular membranes. The structures conformations soluble, overexpressed, purified proteins...
Vesicles assembled from folded, globular proteins have potential for functions different traditional lipid or polymeric vesicles. However, they also present challenges in understanding the assembly process and controlling vesicle properties. From detailed investigation of behavior recombinant fusion proteins, this work reports a simple strategy to engineer protein vesicles containing functional, domains. This is achieved through tunable self‐assembly leucine zippers elastin‐like...
Abstract We present a new approach for macromolecular structure determination from multiple particles in electron cryo-tomography (cryo-ET) data sets. Whereas existing subtomogram averaging approaches are based on 3D models, we propose to optimise regularised likelihood target that approximates function of the 2D experimental images. In addition, analogous Bayesian polishing and contrast transfer (CTF) refinement single-particle analysis, describe exploit increased signal-to-noise ratio...
Abstract Monoclonal antibodies (mAbs) against Ebola virus (EBOV) glycoprotein (GP 1,2 ) are the standard of care for disease (EVD). Anti-GP mAbs targeting stalk and membrane proximal external region (MPER) potently neutralize EBOV in vitro protective a mouse model EVD. However, their neutralization mechanism is poorly understood because they target GP epitope that has evaded structural characterization. Using X-ray crystallography cryo-electron tomography mAb 3A6 complexed with its...
Human respiratory syncytial virus (RSV) is a pneumovirus that causes significant public health burden. Cold-like symptoms are most common, however severe RSV can require hospitalization. leading cause infant hospitalization in the United States. Recently, multiple vaccines have become available for at-risk populations addition to prophylactic monoclonal antibody treatments. We sought better characterize structural organization of filamentous virions guide continued development improved and...
Numerous methods have been developed for immunogold labeling of thick, cryo-preserved biological specimens. However, most the are permutations chemical fixation and sample sectioning, which select isolate immunolabeled region interest. We describe a method combining with cryo-electron microscopy (cryo-EM) tomography (cryo-ET) surface proteins intact mammalian cells or glycoproteins assembling budding viruses in context virus-infected cultured on EM grids. In this method, were maintained...
Caulobacter crescentus is a Gram-negative alphaproteobacterium that commonly lives in oligotrophic fresh- and saltwater environments. C. host to many bacteriophages, including ϕCbK ϕCbK-like which require interaction with the bacterial flagellum pilus complexes during adsorption. It thought six paralogs of flagellin gene present are important for bacteriophage evasion. Here, we show deletion specific flagellins can indeed attenuate adsorption efficiency, although no single completely ablates...
Abstract The evolution of SARS-CoV-2 variants with increased fitness has been accompanied by structural changes in the spike (S) proteins, which are major target for adaptive immune response. Single-particle cryo-EM analysis soluble S protein from revealed this adaptation at high resolution. trimers situ on intact virions potential to provide more functionally relevant insights into structure and virion morphology. Here, we characterized B.1, Alpha, Beta, Gamma, Delta, Kappa, Mu...
Abstract Electron microscopy (EM), cryo-electron (cryo-EM), and tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute largest group pathogenic viruses to humans. The purification enveloped from cell culture presents certain challenges. Specifically, inclusion host-membrane-derived vesicles, complete destruction disruption...
Background Red blood cell (RBC) transfusions are a common, life-saving therapy for many patients, but they have also been associated with poor clinical outcomes. We identified unusual, pleomorphic structures in human RBC transfusion units by negative-stain electron microscopy that appeared identical to those previously reported be bacteria healthy samples. The presence of viable, replicating stored could explain outcomes recipients and major implications medicine. Here, we investigated the...
Human hepatitis B virus core protein (HBc) is a structural of the (HBV) and contributes to HBV regulation host-cell transcription. However, mechanisms transcriptional remain poorly characterized. To dissect function HBc, yeast two-hybrid was performed identify HBc-binding proteins, C-terminal BRG1/hBRM-associated factors 200 (BAF200C) identified. Then, existence HBc interactions with BAF200C full-length BAF200 confirmed via co-immunoprecipitation assays in 293T, HepG2 HepG2-NTCP cells....
Cryo-electron microscopy and single particle averaging (SPA) has been used to determine the structure of numerous purified proteins resolutions less than three Angstroms [1].SPA can also be applied larger biological assemblies including icosahedral helical virions.In fact, large size global symmetry these particles aid in obtaining high-resolution averages [2,3].However, many viruses other are heterogenous lack true making them unsuitable for SPA.Three-dimensional structures targets obtained...