Human species D adenovirus hexon capsid protein mediates cell entry through a direct interaction with CD46

Human adenovirus species D (HAdV-D) types are currently being explored as vaccine vectors for coronavirus disease 2019 (COVID-19) and other severe infectious diseases. The efficacy of such vector-based vaccines depends on functional interactions with receptors on host cells. Adenoviruses of different species are assumed to enter host cells mainly by interactions between the knob domain of the protruding fiber capsid protein and cellular receptors. Using a cell-based receptor-screening assay, we identified CD46 as a receptor for HAdV-D56. The function of CD46 was validated in infection experiments using cells lacking and overexpressing CD46, and by competition infection experiments using soluble CD46. Remarkably, unlike HAdV-B types that engage CD46 through interactions with the knob domain of the fiber protein, HAdV-D types infect host cells through a direct interaction between CD46 and the hexon protein. Soluble hexon proteins (but not fiber knob) inhibited HAdV-D56 infection, and surface plasmon analyses demonstrated that CD46 binds to HAdV-D hexon (but not fiber knob) proteins. Cryoelectron microscopy analysis of the HAdV-D56 virion–CD46 complex confirmed the interaction and showed that CD46 binds to the central cavity of hexon trimers. Finally, soluble CD46 inhibited infection by 16 out of 17 investigated HAdV-D types, suggesting that CD46 is an important receptor for a large group of adenoviruses. In conclusion, this study identifies a noncanonical entry mechanism used by human adenoviruses, which adds to the knowledge of adenovirus biology and can also be useful for development of adenovirus-based vaccine vectors.

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Human Desmoglein-2 and Human CD46 Mediate Human Adenovirus Type 55 Infection, but Human Desmoglein-2 Plays the Major Roles

Journal of Virology ◽

10.1128/jvi.00747-20 ◽

2020 ◽

Vol 94 (17) ◽

Author(s):

Ying Feng ◽

Changhua Yi ◽

Xinglong Liu ◽

Linbing Qu ◽

Wan Su ◽

...

Keyword(s):

Transgenic Mice ◽

Transgenic Mouse ◽

Gene Knockout ◽

Human Adenovirus ◽

Adenovirus Type ◽

Host Cells ◽

Respiratory Pathogen ◽

Cellular Receptors ◽

Fiber Protein ◽

Mouse Lines

ABSTRACT Human adenovirus type 55 (HAdV55) represents an emerging respiratory pathogen and causes severe pneumonia with high fatality in humans. The cellular receptors, which are essential for understanding the infection and pathogenesis of HAdV55, remain unclear. In this study, we found that HAdV55 binding and infection were sharply reduced by disrupting the interaction of viral fiber protein with human desmoglein-2 (hDSG2) but only slightly reduced by disrupting the interaction of viral fiber protein with human CD46 (hCD46). Loss-of-function studies using soluble receptors, blocking antibodies, RNA interference, and gene knockout demonstrated that hDSG2 predominantly mediated HAdV55 infection. Nonpermissive rodent cells became susceptible to HAdV55 infection when hDSG2 or hCD46 was expressed, but hDSG2 mediated more efficient HAd55 infection than hCD46. We generated two transgenic mouse lines that constitutively express either hDSG2 or hCD46. Although nontransgenic mice were resistant to HAdV55 infection, infection with HAdV55 was significantly increased in hDSG2+/+ mice but was much less increased in hCD46+/+ mice. Our findings demonstrate that both hDSG2 and hCD46 are able to mediate HAdV55 infection but hDSG2 plays the major roles. The hDSG2 transgenic mouse can be used as a rodent model for evaluation of HAdV55 vaccine and therapeutics. IMPORTANCE Human adenovirus type 55 (HAdV55) has recently emerged as a highly virulent respiratory pathogen and has been linked to severe and even fatal pneumonia in immunocompetent adults. However, the cellular receptors mediating the entry of HAdV55 into host cells remain unclear, which hinders the establishment of HAdV55-infected animal models and the development of antiviral approaches. In this study, we demonstrated that human desmoglein-2 (hDSG2) plays the major roles during HAdV55 infection. Human CD46 (hCD46) could also mediate the infection of HAdV55, but the efficiency was much lower than for hDSG2. We generated two transgenic mouse lines that express either hDSG2 or hCD46, both of which enabled HAd55 infection in otherwise nontransgenic mice. hDSG2 transgenic mice enabled more efficient HAdV55 infection than hCD46 transgenic mice. Our study adds to our understanding of HAdV55 infection and provides an animal model for evaluating HAdV55 vaccines and therapeutics.

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Sialic Acid-Containing Glycans as Cellular Receptors for Ocular Human Adenoviruses: Implications for Tropism and Treatment

Viruses ◽

10.3390/v11050395 ◽

2019 ◽

Vol 11 (5) ◽

pp. 395 ◽

Cited By ~ 12

Author(s):

Naresh Chandra ◽

Lars Frängsmyr ◽

Sophie Imhof ◽

Rémi Caraballo ◽

Mikael Elofsson ◽

...

Keyword(s):

Sialic Acid ◽

Direct Interaction ◽

Human Adenovirus ◽

Adenovirus Type ◽

Epidemic Keratoconjunctivitis ◽

Surface Plasmon Resonance Analysis ◽

Cellular Receptors ◽

Human Adenoviruses ◽

Fiber Protein ◽

Type 3

Human adenoviruses (HAdV) are the most common cause of ocular infections. Species B human adenovirus type 3 (HAdV-B3) causes pharyngoconjunctival fever (PCF), whereas HAdV-D8, -D37, and -D64 cause epidemic keratoconjunctivitis (EKC). Recently, HAdV-D53, -D54, and -D56 emerged as new EKC-causing agents. HAdV-E4 is associated with both PCF and EKC. We have previously demonstrated that HAdV-D37 uses sialic acid (SA)-containing glycans as cellular receptors on human corneal epithelial (HCE) cells, and the virus interaction with SA is mediated by the knob domain of the viral fiber protein. Here, by means of cell-based assays and using neuraminidase (a SA-cleaving enzyme), we investigated whether ocular HAdVs other than HAdV-D37 also use SA-containing glycans as receptors on HCE cells. We found that HAdV-E4 and -D56 infect HCE cells independent of SAs, whereas HAdV-D53 and -D64 use SAs as cellular receptors. HAdV-D8 and -D54 fiber knobs also bound to cell-surface SAs. Surprisingly, HCE cells were found resistant to HAdV-B3 infection. We also demonstrated that the SA-based molecule i.e., ME0462, designed to bind to SA-binding sites on the HAdV-D37 fiber knob, efficiently prevents binding and infection of several EKC-causing HAdVs. Surface plasmon resonance analysis confirmed a direct interaction between ME0462 and fiber knobs. Altogether, we demonstrate that SA-containing glycans serve as receptors for multiple EKC-causing HAdVs, and, that SA-based compound function as a broad-spectrum antiviral against known and emerging EKC-causing HAdVs.

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α4β1 Integrin Acts as a Cell Receptor for Murine Polyomavirus at the Postattachment Level

Journal of Virology ◽

10.1128/jvi.77.7.3913-3921.2003 ◽

2003 ◽

Vol 77 (7) ◽

pp. 3913-3921 ◽

Cited By ~ 39

Author(s):

Maddalena Caruso ◽

Laura Belloni ◽

Olga Sthandier ◽

Paolo Amati ◽

Marie-Isabelle Garcia

Keyword(s):

Capsid Protein ◽

Cell Receptor ◽

Host Cells ◽

Virus Binding ◽

3T3 Fibroblasts ◽

Virus Adsorption ◽

A Cell ◽

Direct Binding ◽

Murine Polyomavirus ◽

Capsid Protein Vp1

ABSTRACT The initial interaction of murine polyomavirus (Py) with host cells occurs through direct binding of the major capsid protein VP1 with cell membrane molecules containing terminal sialic acids; however, these Py receptor molecules have not yet been identified. Analysis of the capsid protein primary sequences of all murine strains revealed the presence of integrin ligand motifs in the DE and EF loops of VP1 (LDV and DLXXL, respectively) and at the N terminus of VP2 (DGE). We show that infectivity of the Py A2 strain in mouse Swiss 3T3 fibroblasts is significantly reduced only in the presence of natural integrin ligands carrying an LDV motif or antibodies directed against the α4 and β1 integrin subunits. Furthermore, we demonstrate that expression of the α4 subunit in the α4-deficient BALB/c 3T3 cells increases viral infectivity. Addition of α4 function-blocking antibodies, prior to or after virus adsorption, blocks this increased infectivity without affecting virus binding to cells. Taken together, these data indicate that expression of α4 integrin enhances permissivity to Py, probably by acting as one of the postattachment receptors.

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Nutrient-regulated gene expression in eukaryotes

Biochemical Society Symposium ◽

10.1042/bss0730085 ◽

2006 ◽

Vol 73 ◽

pp. 85-96 ◽

Cited By ~ 8

Author(s):

Richard J. Reece ◽

Laila Beynon ◽

Stacey Holden ◽

Amanda D. Hughes ◽

Karine Rébora ◽

...

Keyword(s):

Gene Expression ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Control ◽

Direct Interaction ◽

Signalling Pathways ◽

A Cell ◽

One Step ◽

Higher Eukaryotes ◽

Regulated Gene Expression

The recognition of changes in environmental conditions, and the ability to adapt to these changes, is essential for the viability of cells. There are numerous well characterized systems by which the presence or absence of an individual metabolite may be recognized by a cell. However, the recognition of a metabolite is just one step in a process that often results in changes in the expression of whole sets of genes required to respond to that metabolite. In higher eukaryotes, the signalling pathway between metabolite recognition and transcriptional control can be complex. Recent evidence from the relatively simple eukaryote yeast suggests that complex signalling pathways may be circumvented through the direct interaction between individual metabolites and regulators of RNA polymerase II-mediated transcription. Biochemical and structural analyses are beginning to unravel these elegant genetic control elements.

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Management of Herpesvirus Infections in Hematopoietic Cell Transplant Recipients

Transplantology ◽

10.3390/transplantology2010002 ◽

2021 ◽

Vol 2 (1) ◽

pp. 8-21

Author(s):

Jan Styczynski

Keyword(s):

Cell Transplantation ◽

Hematopoietic Cell Transplantation ◽

Cellular Therapy ◽

Hematopoietic Cell ◽

Host Cells ◽

Preventive Strategy ◽

Cell Transplant ◽

Hematopoietic Cell Transplant ◽

Screening Assay ◽

Specific Level

Following primary infection, herpesviruses establish latency in infected individuals in the host cells and may reactivate upon external stimuli and during periods of immunosuppression. The objective of this paper was to the present current strategies on preventive and therapeutic management of infections with herpesviruses in recipients of hematopoietic cell transplantation. Strategies of antiviral management include prophylaxis, pre-emptive treatment and targeted treatment. Empirical therapy is not used in antiviral strategies. Prophylaxis can be done at universal (preventive strategy) and specific level. Universal prophylaxis includes non-pharmacologic methods of prevention of infection or reactivation. Risk-adapted specific prophylaxis includes use of specific antivirals or cellular therapy or other specific methods in order to prevent specific infection, in high-risk groups. Pre-emptive therapy means use of therapeutic approaches in asymptomatic infection, detected by a screening assay. Targeted therapy is used in established specific viral end-organ infections. The following sections of the paper refer to prophylaxis and treatment strategies, respectively, against CMV, EBV, HSV, VZV, HHV-6, HHV-7, and HHV-8 after allogeneic hematopoietic cell transplantation.

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Human adenovirus serotype 3 fiber protein. Comparison of native and recombinant proteins

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)67887-7 ◽

1991 ◽

Vol 266 (6) ◽

pp. 3961-3967

Author(s):

C Albiges-Rizo ◽

A Barge ◽

R W Ruigrok ◽

P A Timmins ◽

J Chroboczek

Keyword(s):

Recombinant Proteins ◽

Human Adenovirus ◽

Adenovirus Serotype ◽

Fiber Protein

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Characterization of functional domains of the tenascin-R (restrictin) polypeptide: cell attachment site, binding with F11, and enhancement of F11-mediated neurite outgrowth by tenascin-R.

The Journal of Cell Biology ◽

10.1083/jcb.130.2.473 ◽

1995 ◽

Vol 130 (2) ◽

pp. 473-484 ◽

Cited By ~ 46

Author(s):

U Nörenberg ◽

M Hubert ◽

T Brümmendorf ◽

A Tárnok ◽

F G Rathjen

Keyword(s):

Neurite Outgrowth ◽

Cell Attachment ◽

Attachment Site ◽

Direct Interaction ◽

Bacterial Cells ◽

Fibronectin Type Iii ◽

Attachment Region ◽

A Cell ◽

Function Relationship

The extracellular matrix glycoprotein tenascin-R (TN-R) is a multidomain protein implicated in neural cell adhesion. To analyze the structure-function relationship of the different domains of TN-R, several recombinant TN-R fragments were expressed in bacterial cells. Two distinct binding regions were localized on the TN-R polypeptide: a region binding the axon-associated immunoglobulin (Ig)-like F11 protein and a cell attachment site. The binding region of the glycosylphosphatidylinositol (GPI)-anchored F11 was allocated to the second and third fibronectin type III (FNIII)-like domain within TN-R. By using a mutant polypeptide of F11 containing only Ig-like domains, a direct interaction between the Ig-like domains of F11 and FNIII-like domains 2-3 of TN-R was demonstrated. The interaction of TN-R with F11 in in vitro cultures enhanced F11-mediated neurite outgrowth, suggesting that the combined action of F11 and TN-R might be of regulatory influence on axon extension. A cell attachment region was identified in the FNIII-like domain eight of TN-R by domain-specific antibodies and fusion constructs. This site is distinct from the F11 binding site within TN-R.

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Identification of new complement inhibitors using a cell imaging-based, high-throughput screening assay

Molecular Immunology ◽

10.1016/j.molimm.2018.06.145 ◽

2018 ◽

Vol 102 ◽

pp. 184

Author(s):

Feng Lin ◽

Lingjun Zhang ◽

Yuriy Fedorov ◽

Drew J. Adams

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Cell Imaging ◽

Screening Assay ◽

Complement Inhibitors ◽

High Throughput Screening Assay ◽

A Cell

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Calicivirus VP2 forms a portal to mediate endosome escape

10.1101/397901 ◽

2018 ◽

Cited By ~ 2

Author(s):

Michaela Conley ◽

Marion McElwee ◽

Liyana Azmi ◽

Mads Gabrielsen ◽

Olwyn Byron ◽

...

Keyword(s):

Capsid Protein ◽

Conformational Changes ◽

Structural Changes ◽

Host Cells ◽

Major Step ◽

Endosomal Membrane ◽

Endosome Escape ◽

Animal Pathogens ◽

Capsid Shell ◽

Capsid Protein Vp1

AbstractTo 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. TheCaliciviridaeinclude 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 theCaliciviridaeand icosahedral RNA containing viruses in general.

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