Regulation of the initial stages of herpes simplex virus lytic infection and reactivation from latency
Lead Research Organisation:
MRC Virology Unit
Abstract
Herpes simplex virus type 1 (HSV-1) is one of several herpesviruses that infect a high proportion of the human population. Although these viruses do not generally cause dangerous diseases, herpesvirus infections can become serious or even fatal in the newborn and those with damaged immune systems. Genital herpes simplex virus infections are one of the most common sexually transmitted diseases. Herpesviruses are common because after an initial infection they attain a latent state and are carried by the individual for life. As there are no treatments to eradicate latent virus, an understanding of how herpesviruses attain, maintain and reactivate from latency is necessary for improved treatments for herpesvirus diseases. Current research aims to elucidate the molecular mechanisms by which lytic and latent HSV-1 infection are regulated. HSV-1 encodes regulatory proteins that interact with cellular proteins and control systems, either interfering with or hijacking them to serve the needs of the virus. An understanding of these processes will reveal new possibilities for interfering with virus growth and reactivation from latency. Our results will contribute to improved understanding of processes that are important for normal and cancerous cells. The work is laboratory-based and involves biochemistry, molecular biology, microscopy and infection of cultured cells.
Technical Summary
There are several herpesviruses that infect a high proportion of the human population. Although these viruses do not generally cause dangerous diseases, in certain groups of people, particularly the newborn and those with damaged immune systems, herpesvirus infections can become serious or even fatal. Herpesviruses are common because after an initial episode of infection, they attain a latent state so that they are carried by the infected individual for life. As there are no treatments to eradicate latent virus, an understanding of the molecular mechanisms by which herpesviruses attain, maintain and reactivate from latency is necessary for improved treatments for herpesvirus diseases. The balance between latent and lytic herpesvirus infections is governed by both viral and cellular factors, and by antiviral mechanisms that include not only the various arms of the immune defence but also mechanisms that operate via constitutively expressed proteins at the cellular level. This latter phenomenon, which is a relatively recent concept, is known as intrinsic immunity, intrinsic antiviral defence, or intrinsic resistance.
The principle aim of this programme is to decipher the mechanisms of action of herpes simplex virus type 1 (HSV-1) regulatory protein ICP0, which is required for efficient initiation of lytic infection and reactivation of latent virus. The main role of ICP0 is in counteracting intrinsic antiviral defence. We have shown that ICP0 is a ubiquitin E3 ligase that targets cellular intrinsic resistance proteins for destruction by the proteasome pathway. Current research aims to define the mechanisms of substrate targeting by ICP0 and to correlate these activities with the role of ICP0 in the regulation of viral infection. Our approaches utilise cultured cell systems of virus infection.
A major target of ICP0 is the cellular protein PML, a component of nuclear sub-structures known as PML Nuclear Bodies (PML NBs). PML and other PML NB proteins are rapidly recruited to sites associated with infecting HSV-1 genomes, a process that represents an intrinsic cellular response that represses viral gene expression. An important role of ICP0 is to inactivate this response to allow efficient infection. We have determined the molecular basis of the recruitment of PML to HSV-1 genomes and we have found that this mechanism underlies the recruitment of a number of other cellular proteins. We are extending these studies to the regulatory proteins of other herpesviruses, in particular human cytomegalovirus, to determine how their functions and mechanisms of action relate to those of ICP0. The results are expected to increase understanding of processes common to several DNA viral infections. Since components of PML NBs have been implicated in many important cellular control pathways, our experiments are likely to contribute to many aspects of cell biology research.
The principle aim of this programme is to decipher the mechanisms of action of herpes simplex virus type 1 (HSV-1) regulatory protein ICP0, which is required for efficient initiation of lytic infection and reactivation of latent virus. The main role of ICP0 is in counteracting intrinsic antiviral defence. We have shown that ICP0 is a ubiquitin E3 ligase that targets cellular intrinsic resistance proteins for destruction by the proteasome pathway. Current research aims to define the mechanisms of substrate targeting by ICP0 and to correlate these activities with the role of ICP0 in the regulation of viral infection. Our approaches utilise cultured cell systems of virus infection.
A major target of ICP0 is the cellular protein PML, a component of nuclear sub-structures known as PML Nuclear Bodies (PML NBs). PML and other PML NB proteins are rapidly recruited to sites associated with infecting HSV-1 genomes, a process that represents an intrinsic cellular response that represses viral gene expression. An important role of ICP0 is to inactivate this response to allow efficient infection. We have determined the molecular basis of the recruitment of PML to HSV-1 genomes and we have found that this mechanism underlies the recruitment of a number of other cellular proteins. We are extending these studies to the regulatory proteins of other herpesviruses, in particular human cytomegalovirus, to determine how their functions and mechanisms of action relate to those of ICP0. The results are expected to increase understanding of processes common to several DNA viral infections. Since components of PML NBs have been implicated in many important cellular control pathways, our experiments are likely to contribute to many aspects of cell biology research.
Organisations
- MRC Virology Unit, United Kingdom (Lead Research Organisation)
- University of Oxford, United Kingdom (Collaboration)
- University of St Andrews, United Kingdom (Collaboration)
- Friedrich-Alexander University (Collaboration)
- Wellcome Trust, LONDON (Collaboration)
- Nantes University, France (Collaboration)
- Salk Institute for Biological Studies (Collaboration)
- Leibniz Association (Collaboration)
- University of Kansas (Collaboration)
- King's College London, United Kingdom (Collaboration)
People |
ORCID iD |
Roger David Everett (Principal Investigator) |
Publications



Boutell C
(2011)
A viral ubiquitin ligase has substrate preferential SUMO targeted ubiquitin ligase activity that counteracts intrinsic antiviral defence.
in PLoS pathogens

Boutell C
(2008)
Herpes simplex virus type 1 ICP0 phosphorylation mutants impair the E3 ubiquitin ligase activity of ICP0 in a cell type-dependent manner.
in Journal of virology

Boutell C
(2013)
Regulation of alphaherpesvirus infections by the ICP0 family of proteins
in Journal of General Virology

Cuchet D
(2011)
PML isoforms I and II participate in PML-dependent restriction of HSV-1 replication.
in Journal of cell science

Cuchet-Lourenço D
(2013)
The viral ubiquitin ligase ICP0 is neither sufficient nor necessary for degradation of the cellular DNA sensor IFI16 during herpes simplex virus 1 infection.
in Journal of virology

Cuchet-Lourenço D
(2012)
Herpes simplex virus 1 ubiquitin ligase ICP0 interacts with PML isoform I and induces its SUMO-independent degradation.
in Journal of virology

Cuchet-Lourenço D
(2011)
SUMO pathway dependent recruitment of cellular repressors to herpes simplex virus type 1 genomes.
in PLoS pathogens

Everett RD
(2010)
Comparison of the biological and biochemical activities of several members of the alphaherpesvirus ICP0 family of proteins.
in Journal of virology
Description | EC Marie Curie Fellowship Award |
Amount | £154,880 (GBP) |
Funding ID | PIEF-GA-2009- 251948 |
Organisation | Marie Sklodowska-Curie Actions |
Sector | Charity/Non Profit |
Country | Global |
Start | 03/2010 |
End | 03/2012 |
Description | EC SME-STREP contract FP6-037517 'TargetHerpes' |
Amount | £210,000 (GBP) |
Funding ID | LSHG-CT-2006- 037517 |
Organisation | Sixth Framework Programme (FP6) |
Sector | Public |
Country | European Union (EU) |
Start | 01/2007 |
End | 12/2009 |
Description | MRCT Development Gap Funding |
Amount | £128,000 (GBP) |
Organisation | MRC-Technology |
Sector | Private |
Country | United Kingdom |
Start |
Title | Viruses, plasmids, antibodies and cell lines. |
Description | Viruses, plasmids, antibodies and cell lines. We receive many tens of requests for various research tools every year, comprising literally hundreds of items. |
Type Of Material | Technology assay or reagent |
Year Produced | 2006 |
Provided To Others? | Yes |
Impact | Many of our research tools are used by others in their subsequent publications. Far too many to list. |
Description | AAV ICP0 reactivation |
Organisation | University of Nantes |
Department | INSERM U649 (Gene Therapy Laboratory) |
Country | France |
Sector | Academic/University |
PI Contribution | Most of the experimentation |
Collaborator Contribution | Provision of research reagents |
Impact | PMID 16537633 |
Description | BRCA1 study |
Organisation | King's College London |
Department | School of Medicine KCL |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | we did some specialist experimentation and provided advice |
Collaborator Contribution | they did much of the experimentation |
Impact | PMID 16403807 |
Description | CD83 study |
Organisation | Friedrich-Alexander University Erlangen-Nuremberg |
Department | Department of Dermatology |
Country | Germany |
Sector | Academic/University |
PI Contribution | advice and provision of reagents |
Collaborator Contribution | experimental work |
Impact | PMID 17428858 |
Description | HepaRG PML and Sp100 study |
Organisation | Leibniz Association |
Department | Heinrich Pette Institute, Leibniz Institute for Experimental Virology |
Country | Germany |
Sector | Academic/University |
PI Contribution | most of the experimentation |
Collaborator Contribution | provision of cell lines |
Impact | PMID 18160441 |
Start Year | 2006 |
Description | ICP0 and DNA repair |
Organisation | Salk Institute for Biological Studies |
Department | Laboratory of Genetics Salk |
Country | United States |
Sector | Academic/University |
PI Contribution | Provision and exchange of reagents |
Collaborator Contribution | Provision and exchange of reagents |
Impact | PMID 20075863 PMID 21698222 |
Start Year | 2008 |
Description | ICP0 phosphorylation study |
Organisation | University of Kansas |
Department | Department of Molecular Biosciences |
Country | United States |
Sector | Academic/University |
PI Contribution | most of the experimentation |
Collaborator Contribution | provision of research reagents and advice |
Impact | PMID 18715910 |
Description | PML depletion study |
Organisation | Friedrich-Alexander University Erlangen-Nuremberg |
Department | Institute of Clinical and Molecular Virology |
Country | Germany |
Sector | Academic/University |
PI Contribution | most of the experimentation |
Collaborator Contribution | provision of research reagents |
Impact | PMID 16873256 |
Description | STAT1 and IRF3 study |
Organisation | University of St Andrews |
Department | Centre for Biomolecular Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | most of the experimentation |
Collaborator Contribution | provision of research reagents |
Impact | PMID 18579584 |
Start Year | 2007 |
Description | SUMO proteome study |
Organisation | Wellcome Trust |
Department | Wellcome Trust Centre for Gene Regulation and Expression |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Design of study, experimentation, data analysis |
Collaborator Contribution | Design of study, experimentation, data analysis |
Impact | Publications will ensue in due course |
Start Year | 2012 |
Description | proteomics study |
Organisation | University of Oxford |
Department | Department of Biochemistry |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | provision of samples, and later confirmatory analysis of results from a proteomics screen |
Collaborator Contribution | They did a considerable amount of practical work and analysis |
Impact | PMID 19670248 |
Description | MRC Virology Unit Schools day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | An event in which pupils from several local schools receive instruction, demonstration and information on the work of the MRC Virology Unit and general issues in virology. enhanced student interest in virology |
Year(s) Of Engagement Activity | 2006,2007,2008 |