Targeting the ATP-dependent formation of herpesvirus ribonucleoprotein particle assembly as an antiviral approach

Schumann, Sophie; Jackson, Brian R.; Yule, Ian A.; Whitehead, Steven K; Revill, Charlotte; Foster, Richard; Whitehouse, Adrian

doi:10.1038/nmicrobiol.2016.201

Cited by 43 publications

(42 citation statements)

References 55 publications

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“…ADAM 10 specific inhibitor, GI254023X and ADAM 10/17 dual inhibitor, TAPI-2 where purchased from TOCRIS and Merck Millipore, respectively. Cell toxicity was measured using a MTS-based CellTiter 96 AqueousOne Solution Proliferation assay (Promega), as previously described [ 101 ].…”

Section: Methodsmentioning

confidence: 99%

Cellular sheddases are induced by Merkel cell polyomavirus small tumour antigen to mediate cell dissociation and invasiveness

et al. 2018

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Merkel cell carcinoma (MCC) is an aggressive skin cancer with a high propensity for recurrence and metastasis. Merkel cell polyomavirus (MCPyV) is recognised as the causative factor in the majority of MCC cases. The MCPyV small tumour antigen (ST) is considered to be the main viral transforming factor, however potential mechanisms linking ST expression to the highly metastatic nature of MCC are yet to be fully elucidated. Metastasis is a complex process, with several discrete steps required for the formation of secondary tumour sites. One essential trait that underpins the ability of cancer cells to metastasise is how they interact with adjoining tumour cells and the surrounding extracellular matrix. Here we demonstrate that MCPyV ST expression disrupts the integrity of cell-cell junctions, thereby enhancing cell dissociation and implicate the cellular sheddases, A disintegrin and metalloproteinase (ADAM) 10 and 17 proteins in this process. Inhibition of ADAM 10 and 17 activity reduced MCPyV ST-induced cell dissociation and motility, attributing their function as critical to the MCPyV-induced metastatic processes. Consistent with these data, we confirm that ADAM 10 and 17 are upregulated in MCPyV-positive primary MCC tumours. These novel findings implicate cellular sheddases as key host cell factors contributing to virus-mediated cellular transformation and metastasis. Notably, ADAM protein expression may be a novel biomarker of MCC prognosis and given the current interest in cellular sheddase inhibitors for cancer therapeutics, it highlights ADAM 10 and 17 activity as a novel opportunity for targeted interventions for disseminated MCC.

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Section: Methodsmentioning

confidence: 99%

Cellular sheddases are induced by Merkel cell polyomavirus small tumour antigen to mediate cell dissociation and invasiveness

et al. 2018

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“…The ability of ICP27 and ORF57 to interact with different cellular mRNA export factors allows the virus to exploit the inherent redundancy within their hosts and improve the efficiency of viral mRNA export. Targeting these multiple mRNA export factor interactions as part of an antiviral therapy would be challenging; however the inhibition of UAP56 catalysis provides a pinch point to block TREX assembly and a more credible method for combating herpesvirus lytic infection 59 . The data and binding motifs identified in the current study will aid the exploration and improve our understanding of the complex mechanistic details of these multi-protein, multi-interaction assemblies that facilitate mRNA maturation.…”

Section: Discussionmentioning

confidence: 99%

Overlapping motifs on the herpes viral proteins ICP27 and ORF57 mediate interactions with the mRNA export adaptors ALYREF and UIF

Tunnicliffe

Tian

Storer

et al. 2018

Sci Rep

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The TREX complex mediates the passage of bulk cellular mRNA export to the nuclear export factor TAP/NXF1 via the export adaptors ALYREF or UIF, which appear to act in a redundant manner. TREX complex recruitment to nascent RNA is coupled with 5′ capping, splicing and polyadenylation. Therefore to facilitate expression from their intronless genes, herpes viruses have evolved a mechanism to circumvent these cellular controls. Central to this process is a protein from the conserved ICP27 family, which binds viral transcripts and cellular TREX complex components including ALYREF. Here we have identified a novel interaction between HSV-1 ICP27 and an N-terminal domain of UIF in vivo, and used NMR spectroscopy to locate the UIF binding site within an intrinsically disordered region of ICP27. We also characterized the interaction sites of the ICP27 homolog ORF57 from KSHV with UIF and ALYREF using NMR, revealing previously unidentified binding motifs. In both ORF57 and ICP27 the interaction sites for ALYREF and UIF partially overlap, suggestive of mutually exclusive binding. The data provide a map of the binding sites responsible for promoting herpes virus mRNA export, enabling future studies to accurately probe these interactions and reveal the functional consequences for UIF and ALYREF redundancy.

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“…Medium was then aspirated and replaced with pre-warmed complete DMEM (containing inhibitors if applicable) to synchronise infection. Cells were fixed 10 hours post infection and immunofluorescence performed as previously described (64). VP2/3 specific antibodies were used to detect exposed minor capsid proteins, with calnexin antibodies used to visualise proximity to the ER.…”

Section: Methodsmentioning

confidence: 99%

A requirement for Potassium and Calcium Channels during the Endosomal Trafficking of Polyomavirus Virions

Dobson

Mankouri

Whitehouse

2019

Preprint

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19Following internalisation viruses have to escape the endocytic pathway and deliver their 20 genomes to initiate replication. Members of the Polyomaviridae transit through the 21 endolysosomal network and through the endoplasmic reticulum (ER), from which heavily 22 degraded capsids escape into the cytoplasm prior to nuclear entry. Acidification of 23 endosomes and ER entry are essential in the lifecycle of polyomaviruses, however many 24 mechanistic requirements are yet to be elucidated. Alteration of endocytic pH relies upon the 25 activity of ion channels. Using two polyomaviruses with differing capsid architecture, namely 26 Simian virus 40 (SV40) and Merkel cell polyomavirus (MCPyV), we firstly describe methods 27 to rapidly quantify infection using an IncuCyte ZOOM instrument, prior to investigating the 28 role of K + and Ca 2+ channels during early stages of infection. Broad spectrum inhibitors 29 identified that MCPyV, but not SV40, is sensitive to K + channel modulation. In contrast, both 30 viruses are restricted by the broad spectrum Ca 2+ channel inhibitor verapamil, however 31 specific targeting of transient or long lasting Ca 2+ channel subfamilies had no detrimental 32 effect. Further investigation revealed that tetrandrine blockage of two-pore channels (TPCs), 33the activity of which is essential for endolysosomal-ER fusion, ablates infectivity of both 34 MCPyV and SV40 by preventing disassembly of the capsid, which is required for the 35 exposure of minor capsid protein nuclear signals necessary for nuclear transport. This study 36 therefore identifies a novel target to restrict the entry of polyomaviruses. 37 IMPORTANCE 38Polyomaviruses establish ubiquitous, asymptomatic infection in their host. However, in the 39 immunocompromised these viruses can cause a range of potentially fatal diseases. Through 40 the use of SV40 and MCPyV, two polyomaviruses with different capsid organisation, we 41 have investigated the role of ion channels during infection. Here, we show that Ca 2+ channel 42 activity is essential for both polyomaviruses and that MCPyV is also sensitive to K + channel 43 blockage, highlighting new mechanistic requirements of ion channels during polyomavirus 44 infection. In particular, tetrandrine blockage of endolysosomal-ER fusion is highlighted as an 45 essential modulator of both SV40 and MCPyV. Given that the role of ion channels in disease 46 have been well characterised, there is a large panel of clinically available therapeutics that Polyomaviruses (PyVs) are small double stranded DNA viruses that establish persistent 51 infections in their hosts. Whilst infections are generally asymptomatic, PyVs can cause 52 severe disease in the immunosuppressed. Common examples include BKPyV-associated 53 nephropathy and haemorrhagic cystitis, and JCPyV-induced progressive multifocal 54 leukoencephalopathy (PML) (1-3).55 In ~80% of Merkel cell carcinoma (MCC) cases, Merkel cell PyV (MCPyV) infection, clonal 56 integration and UV-mediated mutation of the viral genome occur prior to tumour c...

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Targeting the ATP-dependent formation of herpesvirus ribonucleoprotein particle assembly as an antiviral approach

Cited by 43 publications

References 55 publications

Cellular sheddases are induced by Merkel cell polyomavirus small tumour antigen to mediate cell dissociation and invasiveness

Cellular sheddases are induced by Merkel cell polyomavirus small tumour antigen to mediate cell dissociation and invasiveness

Overlapping motifs on the herpes viral proteins ICP27 and ORF57 mediate interactions with the mRNA export adaptors ALYREF and UIF

A requirement for Potassium and Calcium Channels during the Endosomal Trafficking of Polyomavirus Virions

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