Biology subjects

Conley, M. J.

Publications and source records attributed to Conley, M. J..

4 recordsLinked to original sources

bioRxiv2021

Helical Ordering of Envelope Associated Proteins and Glycoproteins in Respiratory Syncytial Virus Filamentous Virions

Human respiratory syncytial virus (RSV) causes severe respiratory illness in children and the elderly. Treatments for RSV disease are however limited and efforts to produce an effective vaccine have so far been unsuccessful. Understanding RSV virion structure is an important prerequisite for developing interventions to treat or prevent infection but has been challenging because of the fragility of virions propagated in cell culture. Here we show, using cryogenic electron microscopy (cryoEM) and cryogenic electron tomography (cryoET) of RSV particles cultivated directly on transmission electron microscopy (TEM) grids, that there is extensive helical symmetry in RSV filamentous virions. We have calculated a 16 [A] resolution three-dimensional reconstruction of the viral envelope, targeting the matrix protein (M) that forms an endoskeleton below the viral membrane. These data define a helical lattice of M proteins, showing how M is oriented relative to the viral envelope and that helical ordering of viral glycoproteins that stud the viral envelope is coordinated by the M layer. Moreover, the helically ordered viral glycoproteins in RSV filamentous virions cluster in pairs, which may have implications for the conformation of fusion (F) glycoprotein epitopes that are the principal target for vaccine and monoclonal antibody development. We also report the presence, in authentic virus infections, of N-RNA rings packaged within RSV filamentous virions. Overall, the structural data obtained provides molecular insight into the organization of the virion and the mechanism of its assembly.

microbiology↗

bioRxiv2021

The cryo-EM structure of vesivirus 2117 highlights functional variations in entry pathways for viruses in different clades of the vesivirus genus.

Vesivirus 2117 is an adventitious agent that has been responsible for lost productivity in biopharmaceutical production following contamination of Chinese hamster ovary cell cultures in commercial bioreactors. A member of the Caliciviridae, 2117 is classified within the Vesivirus genus in a clade that includes canine and mink caliciviruses but is distinct from the vesicular exanthema of swine clade, which includes the extensively studied feline calicivirus (FCV). We have used cryogenic electron microscopy (cryo-EM) to determine the structure of the capsid of this small, icosahedral, positive-sense RNA containing virus. We show that the outer face of the dimeric capsomeres, which contains the receptor binding site and major immunodominant epitopes in all caliciviruses studied thus far, is quite different from that of FCV. This is a consequence of a 22 amino-acid insertion in the sequence of the FCV major capsid protein that forms a cantilevered arm, which plays an important role in both receptor engagement and undergoes structural rearrangements thought to be important for genome delivery to the cytosol. Our data highlight a potentially important difference in the attachment and entry pathways employed by the different clades of the Vesivirus genus.

microbiology↗

bioRxiv2018

Calicivirus VP2 forms a portal to mediate endosome escape

To initiate the infectious process, many viruses enter their host cells by triggering endocytosis following receptor engagement. The mechanism by which non-enveloped viruses, such as the caliciviruses, escape the endosome is however poorly understood. The Caliciviridae include many important human and animal pathogens, most notably norovirus, the cause of winter vomiting disease. Here we show that VP2, a minor capsid protein encoded by all caliciviruses, forms a large portal assembly at a unique three-fold symmetry axis following receptor engagement. This feature surrounds an open pore in the capsid shell. We hypothesise that the VP2 portal complex is the means by which the virus escapes the endosome, pene-trating the endosomal membrane to release the viral genome into the cytoplasm. Cryogenic electron microscopy (cryoEM) and asymmetric reconstruction were used to investigate structural changes in the capsid of feline calicivirus (FCV) that occur when the virus binds to its cellular receptor junctional adhesion molecule-A (fJAM-A). Near atomic-resolution structures were calculated for the native virion alone and decorated with soluble receptor fragments. We present atomic models of the major capsid protein VP1 in the presence and absence of fJAM-A, revealing the contact interface and conformational changes brought about by the interaction. Furthermore, we have calculated an atomic model of the portal protein VP2 and revealed the structural changes in VP1 that lead to pore formation. While VP2 was known to be critical for the production of infectious virus, its function has been hitherto undetermined. Our finding that VP2 assembles a portal that is likely responsible for endosome escape represents a major step forward in our understanding of both the Caliciviridae and icosahedral RNA containing viruses in general.

microbiology↗

bioRxiv2018

Synergistic Activity of Silver Nanoparticles and Vancomycin Against a Spectrum of Staphylococcus aureus Biofilm Types

Staphylococcus aureus (SA) is an important human pathogen, causing potentially lethal infections of the blood, skin, and lungs. SA is becoming increasingly difficult to treat due to high levels of antibiotic resistance, and new treatments are needed. SA is able to evade antibiotics and immune surveillance through biofilm development. Biofilms are communities of microorganisms that are able to prevent the entry of antimicrobials and immune cells. Biofilms are also involved in SA transmission because biofilms can form on medical devices. In this study, we tested silver nanoparticles and vancomycin for their anti-biofilm effects on SA. We used 10 different SA isolates, representing a spectrum of biofilm-forming ability, and a crystal violet assay to measure biofilm mass. 2g/mL vancomycin treatment resulted in a significant reduction in established SA biofilms in 7/10 isolates, including 4/5 methicillin susceptible SA (MSSA) and 3/5 methicillin-resistant (MRSA) isolates (mean reduction in crystal violet stain of 13.0%; high of 26.5% and low of 0%). Silver nanoparticle treatment of SA biofilms resulted in a significant reduction in 6/10 isolates, including 4/5 MSSA and 2/5 MRSA (mean reduction of 8.7%; high of 21.2% and low of 0%). A combinatorial treatment with silver nanoparticles and vancomycin resulted in significant reductions in 9/10 isolates (mean reduction of 20.8%; high of 39.3% and low of 0%). We conclude that both vancomycin and silver nanoparticle treatment of established tissue culture-based SA biofilms result in significant reductions in biofilm mass, with a combinatorial treatment even more effective than either treatment alone.

microbiology↗