Intrinsic Immunity

Lead Research Organisation: University of Glasgow

Abstract

Dr Chris Boutell, Lead Investigator, MRC-University of Glasgow Centre for Virus Research (CVR), University of Glasgow
The modification of proteins regulates many important aspects of cell biology, including host immunity to infection. Viruses have therefore evolved strategies to utilise or suppress pathways that regulate protein modification to overcome host defences to infection, thereby enhancing their replication, spread, and disease burden. However, these key interactions remain largely undefined at a biochemical level. Work within this programme focuses on two pathways, namely ubiquitin and SUMO (Small Ubiquitin-like MOdifier), which we and others have shown to play an important role in the regulation of host immunity. Our aim is to define the activity of core enzymes that influence the outcome of infection, using herpes simplex virus 1 (HSV-1) as a model DNA virus. These studies will reveal unique insight into the mechanisms that regulate host immunity during DNA virus infection, as well as uncovering fundamental principles that regulate other important areas of cell biology relevant to human disease, for example carcinogenesis. By understanding these principles we aim to develop new targeted approaches for future therapeutic intervention to treat viral infections pertinent to public health.

Technical Summary

The post-translational modification (PTM) of proteins by ubiquitin and ubiquitin-like (UBL) polypeptides regulates many fundamental aspects of cell biology, including multiple facets relating to host immunity. Viruses have therefore evolved mechanisms to utilise or suppress these PTM pathways to overcome host immune defence mechanisms during infection, thereby promoting their replication and spread. Understanding the biochemical interplay between viral and host factors which utilise these pathways will therefore lead to new insights into the regulation of host immunity, processes which restrict the replication of many clinically important viruses. This programme aims to define the biochemistry of key viral host-cell interactions that utilise ubiquitin and SUMO PTM pathways to influence the outcome of host immunity during herpesvirus infection, using HSV-1 as a model system. By defining the biochemistry of these interactions we aim to: significantly advance our understanding of the signalling mechanisms that mediate the cellular restriction of herpesviruses and the establishment of viral latency; uncover fundamental principles relating to the regulation of ubiquitin and SUMO ligases that mediate key aspects of cellular immunity during HSV-1 infection; develop methodologies suitable for the identification of viral ubiquitin ligase inhibitors using ICP0 as a model ligase.
This programme will add significant value to the MRC, CVR, and its associated research partners through the delivery of an interdisciplinary and internationally competitive programme of research. This programme will strengthen the study of ubiquitin and ubiquitin-related pathways within the CVR in other virus systems, including influenza A virus (IAV), Hepatitis C Virus (HCV), and selected arboviruses, through the dissemination of research reagents and expert knowledge. Our goal is to define the functional significance and host-cell regulation of these important PTM pathways during infection, thereby revealing fundamental insight into the biochemical mechanisms that influence the outcome of viral infection and pathogenesis

Publications

10 25 50

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Boutell C (2013) Regulation of alphaherpesvirus infections by the ICP0 family of proteins. in The Journal of general virology

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Busnadiego I (2014) Host and Viral Determinants of Mx2 Antiretroviral Activity in Journal of Virology

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Everett RD (2013) Interplay between viruses and host sumoylation pathways. in Nature reviews. Microbiology

 
Description ASM Conference on Viral Manipulation of Nuclear Processes
Amount $1,000 (USD)
Organisation American Society for Microbiology (ASM) 
Sector Charity/Non Profit
Country United States
Start 11/2014 
End 11/2014
 
Description ASM travel award to 2017 Viral manipulation of nuclear processes meeting
Amount $1,000 (USD)
Organisation American Society for Microbiology (ASM) 
Sector Charity/Non Profit
Country United States
Start 12/2017 
End 12/2017
 
Description BBSRC DTP PhD studentship
Amount £10,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2015 
End 10/2019
 
Description IHW travel award (Dr Ashley Roberts)
Amount $800 (USD)
Organisation International Herpesvirus Workshop 
Sector Multiple
Country Global
Start 07/2018 
End 07/2018
 
Description ISSF Univeristy of Glasgow polyomics funding scheme
Amount £5,000 (GBP)
Organisation University of Glasgow 
Sector Academic/University
Country United Kingdom
Start 11/2016 
 
Description King Abdulaziz University Scholarship scheme
Amount £100,000 (GBP)
Organisation King Abdulaziz University 
Sector Academic/University
Country Saudi Arabia
Start 10/2014 
End 10/2017
 
Description NIH Chromatin Control of Viral Infection
Amount $500 (USD)
Organisation National Institutes of Health (NIH) 
Sector Public
Country United States
Start 09/2014 
End 09/2014
 
Title Transgenic 3D raft culture studies 
Description 3D raft culture is becoming the go to model system for infectious disease research. However, this system has limitations with respect to genetic manipulation studies analysing the role of specific host factors in host immune regulation in response to virus infection. We are optimising cell lines amenable to transgenic manipulation studies and air liquid interface differentiation to generate 3D raft cultures amenable to infectious disease immuno-biology research. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2017 
Provided To Others? No  
Impact Research ongoing 
 
Title in vitro SUMO biochemistry update 
Description The cloning, expression, and purification of various proteins implicated in the regulation of host SUMOylation. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2016 
Provided To Others? Yes  
Impact Conn KL, Wasson P, McFarlane S, Tong L, Brown JR, Grant KG, et al. Novel Role for Protein Inhibitor of Activated STAT 4 (PIAS4) in the Restriction of Herpes Simplex Virus 1 by the Cellular Intrinsic Antiviral Immune Response. Journal of virology. 2016;90(9):4807-26. doi: Brown JR, Conn KL, Wasson P, Charman M, Tong L, Grant K, et al. SUMO Ligase Protein Inhibitor of Activated STAT1 (PIAS1) Is a Constituent Promyelocytic Leukemia Nuclear Body Protein That Contributes to the Intrinsic Antiviral Immune Response to Herpes Simplex Virus 1. Journal of virology. 2016;90(13):5939-52. doi: 10.1128/JVI.00426-16. PubMed PMID: 27099310; PubMed Central PMCID: PMCPMC4907222. 
 
Title in vitro ubiquitin biochemsitry updated 
Description The cloning, expression, and purification of various proteins implicated in ubiquitin biochemistry. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2016 
Provided To Others? Yes  
Impact 1. Lilley, C. E., M. S. Chaurushiya, C. Boutell, S. Landry, J. Suh, S. Panier, R. D. Everett, G. S. Stewart, D. Durocher, and M. D. Weitzman. 2010. A viral E3 ligase targets RNF8 and RNF168 to control histone ubiquitination and DNA damage responses. Embo J 29:943-55. 2. Morris, J. R., C. Boutell, M. Keppler, R. Densham, D. Weekes, A. Alamshah, L. Butler, Y. Galanty, L. Pangon, T. Kiuchi, T. Ng, and E. Solomon. 2009. The SUMO modification pathway is involved in the BRCA1 response to genotoxic stress. Nature 462:886-90. 3. Morris, J. R., L. Pangon, C. Boutell, T. Katagiri, N. H. Keep, and E. Solomon. 2006. Genetic analysis of BRCA1 ubiquitin ligase activity and its relationship to breast cancer susceptibility. Hum Mol Genet 15:599-606. 
 
Title shRNA lentiviral cell lines updated 2016 
Description Primary cell lines that have been transduced with lentiviral vectors expressing short hairpin RNAs (shRNAs) to deplete components of the ubiquitin and SUMO conjugation pathways. 
Type Of Material Cell line 
Year Produced 2016 
Provided To Others? Yes  
Impact Research ongoing. Conn KL, Wasson P, McFarlane S, Tong L, Brown JR, Grant KG, et al. Novel Role for Protein Inhibitor of Activated STAT 4 (PIAS4) in the Restriction of Herpes Simplex Virus 1 by the Cellular Intrinsic Antiviral Immune Response. Journal of virology. 2016;90(9):4807-26. doi: Brown JR, Conn KL, Wasson P, Charman M, Tong L, Grant K, et al. SUMO Ligase Protein Inhibitor of Activated STAT1 (PIAS1) Is a Constituent Promyelocytic Leukemia Nuclear Body Protein That Contributes to the Intrinsic Antiviral Immune Response to Herpes Simplex Virus 1. Journal of virology. 2016;90(13):5939-52. doi: 10.1128/JVI.00426-16. PubMed PMID: 27099310; PubMed Central PMCID: PMCPMC4907222. 
 
Title single genome labelling technologies to study virus host-cell interactions 
Description viral nucleic acid represents a major immune stimulus. However, visualization of viral nucleic acid upon entry to host cells in relation to cellular proteins which mediate immune signalling has been problematic. We have optimized an efficient methodology for labeling viral DNA that has enabled us to detect viral nucleic acid within cells 15 minutes post-infection. This technique will prove extremely valuable in the identification of cellular processes that directly contribute to regulation of host immunity to virus infection. 
Type Of Material Biological samples 
Year Produced 2017 
Provided To Others? Yes  
Impact Alandijany T, Roberts APE, Conn KL, Loney C, McFarlane S, Orr A, et al. Distinct temporal roles for the promyelocytic leukaemia (PML) protein in the sequential regulation of intracellular host immunity to HSV-1 infection. PLoS pathogens. 2018;14(1):e1006769. doi: 10.1371/journal.ppat.1006769. PubMed PMID: 29309427; PubMed Central PMCID: PMCPMC5757968. 
 
Title Transcriptome data of HSV-1 infected HFt cells depleted of HIRA 
Description in vitro model of human fibroblast cells depleted of the histone H3.3 chaperone protein HIRA and left untreated, stimulated with interferon-beta (100IU/ml), or infected with HSV-1 
Type Of Material Database/Collection of data 
Year Produced 2019 
Provided To Others? Yes  
Impact Manuscript accepted for publication 
URL https://www.ebi.ac.uk/ena/data/search?query=PRJEB27501
 
Description Analysing the role of Sp1 to mediate cGAS/IFI16 detection of HSV-1 genomes and induction of host innate immune defences 
Organisation University of Ulm
Country Germany 
Sector Academic/University 
PI Contribution Investigation into the role of the transcription factor Sp1 in the regulation and sensing of viral nucleic acid acid in response to HSV-1 infection and induction of host innate immune defences.
Collaborator Contribution Access to preliminary data on the role of Sp1 in the regulation of innate immunity in response to HIV infection.
Impact Multi-disciplinary: HIV, HSV-1, cell biology, host immunology
Start Year 2019
 
Description Astrazeneca research partnership 
Organisation AstraZeneca
Department Research and Development AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Provision of academic support in the development of infectious disease research in the area of autophagy and respiratory disease
Collaborator Contribution Employment of a post-doctoral scientist to investigate autophagy and infectious respiratory disease
Impact no impact yet
Start Year 2017
 
Description Defining the role of SUMOylation during arbovirus infection 
Organisation The Pirbright Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution Defining the biochemical activity of host SUMOylation enzymes expressed in Aedes aegepti.
Collaborator Contribution Cloning of cDNAs that express Aedes aegepti SUMOylation enzymes
Impact Research ongoing. Student selected for national presentation (microbiology society)
Start Year 2014
 
Description Identification of PML-NB host factors by LC-MSMS proteomics 
Organisation University of Pennsylvania
Country United States 
Sector Academic/University 
PI Contribution Provision of materials and reagents for study
Collaborator Contribution Construction of cell lines and LC-MSMS analysis
Impact No outputs yet
Start Year 2017
 
Description Intrinsic immunity to Influenza A Virus 
Organisation University of Glasgow
Department MRC - University of Glasgow Centre for Virus Research
Country United Kingdom 
Sector Academic/University 
PI Contribution Provision or reagents and research support to establish a role for intrinsic immunity against IAV.
Collaborator Contribution Provision or reagents and research support to establish a role for intrinsic immunity against IAV.
Impact We have identfied the TRIM protein 22 to play a role in the cellular restriction of IAV prior to the activation of innate immune signalling, thereby confirming a role for intrinsic immunity in the intracellular restriction of IAV. A manuscript is currently in preparation.
Start Year 2015
 
Description Investigating the role of the histone chaperone protein HIRA in the regulation of host innate immune defences to HSV-1 infection 
Organisation Sanford Burnham Prebys Medical Discovery Institute
PI Contribution Investigation into the role of the H3.3 histone chaperone protein HIRA in the regulation of host innate immune defences in response to HSV-1 infection
Collaborator Contribution Provision of research reagents
Impact Manuscript accepted for publication
Start Year 2018
 
Description Investigating the role of the histone chaperone protein HIRA in the regulation of host innate immune defences to HSV-1 infection 
Organisation Ulster University
Department School of Biomedical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Investigation into the role of the H3.3 histone chaperone protein HIRA in the regulation of host innate immune defences in response to HSV-1 infection
Collaborator Contribution Provision of research reagents
Impact Manuscript accepted for publication
Start Year 2018
 
Description MedImmune industrial student partnership 
Organisation MedImmune
Country United States 
Sector Private 
PI Contribution Investigating the role of intrinsic immunity to restrict the replication of live attenuated influenza viruses (LAIV) that are used as vaccines to protect against influenza virus infection in children. Me and my team provide research support to aid the training of MedImmune-UoG IPP student to investigate the role of intrinsic immunity to restrict LAIV replication and to establish a cost effective cell culture system to support IAV infection and immune-biology studies.
Collaborator Contribution Provision of research reagents (LAIV strains) and financial backing to underpin research studentship
Impact The initiation of this collaboration has led to the provisional contact with other industrial partners, for example BiogelX (https://www.biogelx.com), to establish a viable low cost cell culture system amenable to transgenic manipulation through CRISPR/CAS9 gene editing. Multi-disciplinary: industrial research, vaccine design, molecular virology, gene editing.
Start Year 2019
 
Description Mosquito invertebrate SUMO biology research 
Organisation University of Glasgow
Department MRC - University of Glasgow Centre for Virus Research
Country United Kingdom 
Sector Academic/University 
PI Contribution Provision of material for mosquito SUMO biology research
Collaborator Contribution provision of vector species knowledge and expertise
Impact manuscript in preparation
Start Year 2016
 
Description Proteomic analysis of arbovirus infected invertebrate cells 
Organisation University of Dundee
Country United Kingdom 
Sector Academic/University 
PI Contribution provision of material for proteomic analysis; in vitro biochemical samples, infected invertebrate cells, infected mosquito salivary glands.
Collaborator Contribution LC-MSMS analysis
Impact manuscript in preparation
Start Year 2017
 
Description Visualisation of HSV-1 latency 
Organisation University of Virginia (UVa)
Country United States 
Sector Academic/University 
PI Contribution Provision of research reagents and technical knowledge to the visualisation of latent viral genomes
Collaborator Contribution Provision of neuronal culture systems for studying HSV-1 viral latency and reactivation in vitro
Impact No outputs yet
Start Year 2017
 
Description X-ray crystallography studies of the purified ICP0 RING-finger domains from HSV-1 and VZV 
Organisation York University Toronto
Country Canada 
Sector Academic/University 
PI Contribution Provision of reagents for the purification of RING-domain containing viral E3 ubiquitin ligases expressed during HSV-1 and VZV infection
Collaborator Contribution Purification of recombinant proteins and X-ray crystallography trials
Impact none
Start Year 2015
 
Description 23rd Glasgow Virology Workshop 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact One day scientific symposium to promote awareness about virology research and its impact in our society. over 150 delegates attended with local, national, and international speakers.
Year(s) Of Engagement Activity 2013,2014,2015,2016,2017
URL https://www.gla.ac.uk/researchinstitutes/iii/cvr/events/headline_430158_en.html
 
Description 3 minute thesis presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Study participants or study members
Results and Impact Open forum to discuss research activities in a layman language to engage public sector in scientific research
Year(s) Of Engagement Activity 2018
 
Description February 2017 - Chris Boutell - BBC commentary 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Dr Chris Boutell provided guidance and information about herpes simplex virus for a short item about cold sores on Trust Me I'm a Doctor. The show aired on prime time BBC2 in February 2017. this was also shared on our social media channels to highlight the show to staff and students across the CVR. The post reached 1114 followers with likes and 2 shares on facebook along being shared with >200 followers on Twitter.
Year(s) Of Engagement Activity 2017
URL https://www.facebook.com/centreforvirusresearch
 
Description February 2017 Trust me I'm a Doctor - Dr Chris Boutell 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact In September 2016 Dr Chris Boutell was approached by the BBC and asked to review content for a show that would air on BBC 2 called Trust me I'm a Doctor. The show contained a short segment on herpes virus and Dr Boutell was asked questions and provided commentary on content to ensure accuracy. The show aired on prime time television in February 2017.
Year(s) Of Engagement Activity 2017
URL https://www.facebook.com/centreforvirusresearch
 
Description Glasgow Virology Workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact This annual one day meeting aims to encourage interactions between virologists within Scotland and across the UK. It is usually very well attended (over 200 delegates last year), with groups from Glasgow, Edinburgh, St. Andrews, Dundee, Leeds, as well as other universities from around the UK. We attract a diverse audience, including local biotechnology companies, professional peers, undergraduates, and post-graduate students.
Year(s) Of Engagement Activity 2013,2014,2015,2016,2017
URL http://www.gla.ac.uk/researchinstitutes/iii/cvr/events/headline_430158_en.html
 
Description Kelvingrove Museum Science Weekend 2014 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact We ran our 'Build a virus' and 'One Infection' activities - offering participants the opportunity to meet CVR scientists, learn about the structure of viruses and also key messages related to the transmission of viruses, and control measures including hygiene and vaccination.

Participants were asked to provide feedback on the activities to help us determine how to improve the activity in future. We have been asked to submit a proposal to run an activity during the Glasgow Science Festival in June 2015.
Year(s) Of Engagement Activity 2014
 
Description PCR workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Since 2006, this activity has been delivered to more than 700 Scottish high school students. The teaching resources developed have been adopted by Learning Teaching Scotland to teach higher biology. Workshop has been praised in an HMIE report. Evaluation student feedback is very positive each year.

Our PCR workshop run in partnership with Glasgow Science Centre is a flagship/exemplar activity in high-school education. The PCR workshop has been cited by HMiE as an example of excellence. School pupils from all over Scotland travel to Glasgow to participate.
Year(s) Of Engagement Activity 2013,2014
 
Description PhD student Thamir Alandijany - Saudi International TV Live Interview 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact CVR PhD Student Thamir Alandijany gave a live interview on the Saudi television channel. We don't have a clip available but this was shared on Facebook and it has been included in our PEC report for 2018. Thamir was interviewed about his experience of completing a PhD in Scotland/UK.
Year(s) Of Engagement Activity 2018
URL https://www.facebook.com/centreforvirusresearch/
 
Description Scientific consultant for BBC 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Scientific consultant for a BBC public engagement program explaining the cause of cold sores
Year(s) Of Engagement Activity 2017
 
Description Scout's Science Day - 2014 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 200 local scouts and guides attended a day of science engagement at the University of Glasgow where they had the opportunity to take part in a range of activities. All attendees took part in our 'Build a Virus' activity and meet with CVR scientists. These interactions gave the attendees the opportunity to ask questions and also learn a little more about viruses, their structure, transmission and control through group discussion with individual researchers. The activity was supported by 8 CVR researchers, providing an excellent opportunity for individual public engagement training and development.

The local Scout and Guide leaders are keen to facilitate another event in 2015.
Year(s) Of Engagement Activity 2014
 
Description Thamir Alandijany - Saudi International Television Interview - 23 May 2018 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact PhD student Thamir Alandijany was interviewed on the Saudi Arabia national TV channel - sharing his experience of working and completing his studies at the CVR. Thamir also talked about his recently published article on herpes virology and his experience of winning awards throughout his PhD. No link available.
Year(s) Of Engagement Activity 2018