Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection

ABSTRACT Herpesviruses utilize multiple mechanisms to redirect host proteins for use in viral processes and to avoid recognition and repression by the host. To investigate dynamic interactions between herpes simplex virus type 1 (HSV-1) DNA and viral and host proteins throughout infection, we develo...

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Autores principales:	Jill A. Dembowski, Neal A. DeLuca
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	DNA damage DNA repair ICP4 affinity purification herpes simplex virus iPOND Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9b1e7df7b1cf44aa9febb3ef1fc123f9
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spelling	oai:doaj.org-article:9b1e7df7b1cf44aa9febb3ef1fc123f92021-11-15T16:00:16ZTemporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection10.1128/mBio.01182-182150-7511https://doaj.org/article/9b1e7df7b1cf44aa9febb3ef1fc123f92018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01182-18https://doaj.org/toc/2150-7511ABSTRACT Herpesviruses utilize multiple mechanisms to redirect host proteins for use in viral processes and to avoid recognition and repression by the host. To investigate dynamic interactions between herpes simplex virus type 1 (HSV-1) DNA and viral and host proteins throughout infection, we developed an approach to identify proteins that associate with the infecting viral genome from nuclear entry through packaging. To accomplish this, virus stocks were prepared in the presence of ethynyl-modified nucleotides to enable covalent tagging of viral genomes after infection for analysis of viral genome-protein interactions by imaging or affinity purification. Affinity purification was combined with stable isotope labeling of amino acids in cell culture (SILAC) mass spectrometry to enable the distinction between proteins that were brought into the cell by the virus or expressed within the infected cell before or during infection. We found that input viral DNA progressed within 6 h through four temporal stages where the genomes sequentially (i) interacted with intrinsic antiviral and DNA damage response proteins, (ii) underwent a robust transcriptional switch mediated largely by ICP4, (iii) engaged in replication, repair, and continued transcription, and then (iv) transitioned to a more transcriptionally inert state engaging de novo-synthesized viral structural components while maintaining interactions with replication proteins. Using a combination of genetic, imaging, and proteomic approaches, we provide a new and temporally compressed view of the HSV-1 life cycle based on input genome-proteome dynamics. IMPORTANCE Herpesviruses are highly prevalent and ubiquitous human pathogens. Studies of herpesviruses and other viruses have previously been limited by the ability to directly study events that occur on the viral DNA throughout infection. We present a new powerful approach, which allows for the temporal investigation of viral genome-protein interactions at all phases of infection. This work has integrated many results from previous studies with the discovery of novel factors potentially involved in viral infection that may represent new antiviral targets. In addition, the study provides a new view of the HSV-1 life cycle based on genome-proteome dynamics.Jill A. DembowskiNeal A. DeLucaAmerican Society for MicrobiologyarticleDNA damageDNA repairICP4affinity purificationherpes simplex virusiPONDMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	DNA damage DNA repair ICP4 affinity purification herpes simplex virus iPOND Microbiology QR1-502
spellingShingle	DNA damage DNA repair ICP4 affinity purification herpes simplex virus iPOND Microbiology QR1-502 Jill A. Dembowski Neal A. DeLuca Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
description	ABSTRACT Herpesviruses utilize multiple mechanisms to redirect host proteins for use in viral processes and to avoid recognition and repression by the host. To investigate dynamic interactions between herpes simplex virus type 1 (HSV-1) DNA and viral and host proteins throughout infection, we developed an approach to identify proteins that associate with the infecting viral genome from nuclear entry through packaging. To accomplish this, virus stocks were prepared in the presence of ethynyl-modified nucleotides to enable covalent tagging of viral genomes after infection for analysis of viral genome-protein interactions by imaging or affinity purification. Affinity purification was combined with stable isotope labeling of amino acids in cell culture (SILAC) mass spectrometry to enable the distinction between proteins that were brought into the cell by the virus or expressed within the infected cell before or during infection. We found that input viral DNA progressed within 6 h through four temporal stages where the genomes sequentially (i) interacted with intrinsic antiviral and DNA damage response proteins, (ii) underwent a robust transcriptional switch mediated largely by ICP4, (iii) engaged in replication, repair, and continued transcription, and then (iv) transitioned to a more transcriptionally inert state engaging de novo-synthesized viral structural components while maintaining interactions with replication proteins. Using a combination of genetic, imaging, and proteomic approaches, we provide a new and temporally compressed view of the HSV-1 life cycle based on input genome-proteome dynamics. IMPORTANCE Herpesviruses are highly prevalent and ubiquitous human pathogens. Studies of herpesviruses and other viruses have previously been limited by the ability to directly study events that occur on the viral DNA throughout infection. We present a new powerful approach, which allows for the temporal investigation of viral genome-protein interactions at all phases of infection. This work has integrated many results from previous studies with the discovery of novel factors potentially involved in viral infection that may represent new antiviral targets. In addition, the study provides a new view of the HSV-1 life cycle based on genome-proteome dynamics.
format	article
author	Jill A. Dembowski Neal A. DeLuca
author_facet	Jill A. Dembowski Neal A. DeLuca
author_sort	Jill A. Dembowski
title	Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
title_short	Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
title_full	Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
title_fullStr	Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
title_full_unstemmed	Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection
title_sort	temporal viral genome-protein interactions define distinct stages of productive herpesviral infection
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/9b1e7df7b1cf44aa9febb3ef1fc123f9
work_keys_str_mv	AT jilladembowski temporalviralgenomeproteininteractionsdefinedistinctstagesofproductiveherpesviralinfection AT nealadeluca temporalviralgenomeproteininteractionsdefinedistinctstagesofproductiveherpesviralinfection
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Temporal Viral Genome-Protein Interactions Define Distinct Stages of Productive Herpesviral Infection

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