Temporal Proteomic Analysis of Herpes Simplex Virus 1 Infection Reveals Cell-Surface Remodeling via pUL56-Mediated GOPC Degradation

Soh, Timothy K.; Davies, Colin; Muenzner, Julia; Hunter, Leah M.; Barrow, Henry G.; Connor, Viv; Bouton, Clément R.; Smith, C. L.; Emmott, Edward; Antrobus, Robin; Graham, Stephen C.; Weekes, Michael P.; Crump, Colin M.

doi:10.1016/j.celrep.2020.108235

Cited by 38 publications

(45 citation statements)

References 100 publications

(136 reference statements)

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“…GST pull-down experiments using reagents purified following recombinant expression in Escherichia coli (pUL21 and PP1γ) and HEK293F cells (CERT) confirmed that pUL21 binds directly to both PP1 and CERT (Fig 1C). Published quantitative viromics analysis [27] confirms that the abundance of CERT and all three PP1 catalytic subunit isoforms is unchanged in keratinocytes over the course of HSV-1 infection.…”

Section: Resultsmentioning

confidence: 84%

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

et al. 2021

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The herpes simplex virus (HSV)-1 protein pUL21 is essential for efficient virus replication and dissemination. While pUL21 has been shown to promote multiple steps of virus assembly and spread, the molecular basis of its function remained unclear. Here we identify that pUL21 is a virus-encoded adaptor of protein phosphatase 1 (PP1). pUL21 directs the dephosphorylation of cellular and virus proteins, including components of the viral nuclear egress complex, and we define a conserved non-canonical linear motif in pUL21 that is essential for PP1 recruitment. In vitro evolution experiments reveal that pUL21 antagonises the activity of the virus-encoded kinase pUS3, with growth and spread of pUL21 PP1-binding mutant viruses being restored in adapted strains where pUS3 activity is disrupted. This study shows that virus-directed phosphatase activity is essential for efficient herpesvirus assembly and spread, highlighting the fine balance between kinase and phosphatase activity required for optimal virus replication.

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Section: Resultsmentioning

confidence: 84%

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

et al. 2021

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“…S1 B ). Using multiplexed quantitative proteomic analysis over the whole time course of HSV-1 infection, it has recently been shown that the viral glycoproteins gM and gC follow similar kinetic expression profiles ( 20 ). We demonstrate first by using time-lapse imaging that indeed gM behaves in a similar way to gC, which allows for spatial and temporal correlation with our previous data sets acquired with the timestamp virus ( Fig.…”

Section: Resultsmentioning

confidence: 99%

A fluorescent reporter system enables spatiotemporal analysis of host cell modification during herpes simplex virus-1 replication

Scherer

Manton

Soh

et al. 2021

Journal of Biological Chemistry

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Herpesviruses are large and complex viruses that have a long history of coevolution with their host species. One important factor in the virus–host interaction is the alteration of intracellular morphology during viral replication with critical implications for viral assembly. However, the details of this remodeling event are not well understood, in part because insufficient tools are available to deconstruct this highly heterogeneous process. To provide an accurate and reliable method of investigating the spatiotemporal dynamics of virus-induced changes to cellular architecture, we constructed a dual-fluorescent reporter virus that enabled us to classify four distinct stages in the infection cycle of herpes simplex virus-1 at the single cell level. This timestamping method can accurately track the infection cycle across a wide range of multiplicities of infection. We used high-resolution fluorescence microscopy analysis of cellular structures in live and fixed cells in concert with our reporter virus to generate a detailed and chronological overview of the spatial and temporal reorganization during viral replication. The highly orchestrated and striking relocation of many organelles around the compartments of secondary envelopment during transition from early to late gene expression suggests that the reshaping of these compartments is essential for virus assembly. We furthermore find that accumulation of HSV-1 capsids in the cytoplasm is accompanied by fragmentation of the Golgi apparatus with potential impact on the late steps of viral assembly. We anticipate that in the future similar tools can be systematically applied for the systems-level analysis of intracellular morphology during replication of other viruses.

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“…Immunoblotting for phosphorylated Akt substrates in HSV-1 infected cells reveals additional proteins targeted by pUL21 for PP1-mediated dephosphorylation that have yet to be identified and confirms that pUL21 has a broad specificity that overlaps with, but is not identical to, that of pUS3. In contrast to pUS3, which is expressed immediately after infection and plateaus at around 6 hours post-infection (hpi), pUL21 is a late gene and its abundance increases steadily as the infection progresses (Soh et al, 2020). One could thus hypothesise that pUL21 confers temporal regulation of pUS3-dependent protein phosphorylation, promoting dephosphorylation of specific pUS3 substrates at late times post-infection to ensure coordinated progression of virus replication.…”

Section: Discussionmentioning

confidence: 99%

“…Mass spectrometry analysis was performed by the proteomics facility of the University of Bristol (UK) as described in (Soh et al, 2020). The raw data files were processed using MaxQuant v. 1.5.6.0 (Cox and Mann, 2008).…”

Section: Gfp Affinity Capture and Quantitative Mass Spectrometrymentioning

confidence: 99%

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

Benedyk

Muenzner

Connor

et al. 2021

Preprint

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The herpes simplex virus (HSV)-1 protein pUL21 is essential for efficient virus replication and dissemination. While pUL21 has been shown to promote multiple steps of virus assembly and spread, the molecular basis of its function remained unclear. Here we identify that pUL21 is a virus-encoded adaptor of protein phosphatase 1 (PP1). pUL21 directs the dephosphorylation of cellular and virus proteins, including components of the viral nuclear egress complex, and we define a conserved non-canonical linear motif in pUL21 that is essential for PP1 recruitment. In vitro evolution experiments reveal that pUL21 directly antagonises the activity of the virus-encoded kinase pUS3, with growth and spread of pUL21 PP1-binding mutant viruses being restored when pUS3 activity is disrupted. This study shows that virus-directed phosphatase activity is essential for efficient herpesvirus assembly and spread, highlighting the fine balance between kinase and phosphatase activity required for optimal virus replication.

show abstract

Temporal Proteomic Analysis of Herpes Simplex Virus 1 Infection Reveals Cell-Surface Remodeling via pUL56-Mediated GOPC Degradation

Cited by 38 publications

References 100 publications

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

A fluorescent reporter system enables spatiotemporal analysis of host cell modification during herpes simplex virus-1 replication

pUL21 is a viral phosphatase adaptor that promotes herpes simplex virus replication and spread

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