Dynamic Proteomics of Herpes Simplex Virus Infection

ABSTRACT The cellular response to viral infection is usually studied at the level of cell populations. Currently, it remains an open question whether and to what extent cell-to-cell variability impacts the course of infection. Here we address this by dynamic proteomics—imaging and tracking 400 yello...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Nir Drayman, Omer Karin, Avi Mayo, Tamar Danon, Lev Shapira, Dor Rafael, Anat Zimmer, Anat Bren, Oren Kobiler, Uri Alon
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	cell cycle geminin RFX7 RPAP3 SLTM single-cell infection Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5a469c18db8f4c55bd316f119c67c6ab
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:5a469c18db8f4c55bd316f119c67c6ab
record_format	dspace
spelling	oai:doaj.org-article:5a469c18db8f4c55bd316f119c67c6ab2021-11-15T15:51:56ZDynamic Proteomics of Herpes Simplex Virus Infection10.1128/mBio.01612-172150-7511https://doaj.org/article/5a469c18db8f4c55bd316f119c67c6ab2017-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01612-17https://doaj.org/toc/2150-7511ABSTRACT The cellular response to viral infection is usually studied at the level of cell populations. Currently, it remains an open question whether and to what extent cell-to-cell variability impacts the course of infection. Here we address this by dynamic proteomics—imaging and tracking 400 yellow fluorescent protein (YFP)-tagged host proteins in individual cells infected by herpes simplex virus 1. By quantifying time-lapse fluorescence imaging, we analyze how cell-to-cell variability impacts gene expression from the viral genome. We identify two proteins, RFX7 and geminin, whose levels at the time of infection correlate with successful initiation of gene expression. These proteins are cell cycle markers, and we find that the position in the cell cycle at the time of infection (along with the cell motility and local cell density) can reasonably predict in which individual cells gene expression from the viral genome will commence. We find that the onset of cell division dramatically impacts the progress of infection, with 70% of dividing cells showing no additional gene expression after mitosis. Last, we identify four host proteins that are specifically modulated in infected cells, of which only one has been previously recognized. SUMO2 and RPAP3 levels are rapidly reduced, while SLTM and YTHDC1 are redistributed to form nuclear foci. These modulations are dependent on the expression of ICP0, as shown by infection with two mutant viruses that lack ICP0. Taken together, our results provide experimental validation for the long-held notion that the success of infection is dependent on the state of the host cell at the time of infection. IMPORTANCE High-throughput assays have revolutionized many fields in biology, both by allowing a more global understanding of biological processes and by deciphering rare events in subpopulations. Here we use such an assay, dynamic proteomics, to study viral infection at the single-cell level. We follow tens of thousands of individual cells infected by herpes simplex virus using fluorescence live imaging. Our results link the state of a cell at the time of virus infection with its probability to successfully initiate gene expression from the viral genome. Further, we identified three cellular proteins that were previously unknown to respond to viral infection. We conclude that dynamic proteomics provides a powerful tool to study single-cell differences during viral infection.Nir DraymanOmer KarinAvi MayoTamar DanonLev ShapiraDor RafaelAnat ZimmerAnat BrenOren KobilerUri AlonAmerican Society for Microbiologyarticlecell cyclegemininRFX7RPAP3SLTMsingle-cell infectionMicrobiologyQR1-502ENmBio, Vol 8, Iss 6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	cell cycle geminin RFX7 RPAP3 SLTM single-cell infection Microbiology QR1-502
spellingShingle	cell cycle geminin RFX7 RPAP3 SLTM single-cell infection Microbiology QR1-502 Nir Drayman Omer Karin Avi Mayo Tamar Danon Lev Shapira Dor Rafael Anat Zimmer Anat Bren Oren Kobiler Uri Alon Dynamic Proteomics of Herpes Simplex Virus Infection
description	ABSTRACT The cellular response to viral infection is usually studied at the level of cell populations. Currently, it remains an open question whether and to what extent cell-to-cell variability impacts the course of infection. Here we address this by dynamic proteomics—imaging and tracking 400 yellow fluorescent protein (YFP)-tagged host proteins in individual cells infected by herpes simplex virus 1. By quantifying time-lapse fluorescence imaging, we analyze how cell-to-cell variability impacts gene expression from the viral genome. We identify two proteins, RFX7 and geminin, whose levels at the time of infection correlate with successful initiation of gene expression. These proteins are cell cycle markers, and we find that the position in the cell cycle at the time of infection (along with the cell motility and local cell density) can reasonably predict in which individual cells gene expression from the viral genome will commence. We find that the onset of cell division dramatically impacts the progress of infection, with 70% of dividing cells showing no additional gene expression after mitosis. Last, we identify four host proteins that are specifically modulated in infected cells, of which only one has been previously recognized. SUMO2 and RPAP3 levels are rapidly reduced, while SLTM and YTHDC1 are redistributed to form nuclear foci. These modulations are dependent on the expression of ICP0, as shown by infection with two mutant viruses that lack ICP0. Taken together, our results provide experimental validation for the long-held notion that the success of infection is dependent on the state of the host cell at the time of infection. IMPORTANCE High-throughput assays have revolutionized many fields in biology, both by allowing a more global understanding of biological processes and by deciphering rare events in subpopulations. Here we use such an assay, dynamic proteomics, to study viral infection at the single-cell level. We follow tens of thousands of individual cells infected by herpes simplex virus using fluorescence live imaging. Our results link the state of a cell at the time of virus infection with its probability to successfully initiate gene expression from the viral genome. Further, we identified three cellular proteins that were previously unknown to respond to viral infection. We conclude that dynamic proteomics provides a powerful tool to study single-cell differences during viral infection.
format	article
author	Nir Drayman Omer Karin Avi Mayo Tamar Danon Lev Shapira Dor Rafael Anat Zimmer Anat Bren Oren Kobiler Uri Alon
author_facet	Nir Drayman Omer Karin Avi Mayo Tamar Danon Lev Shapira Dor Rafael Anat Zimmer Anat Bren Oren Kobiler Uri Alon
author_sort	Nir Drayman
title	Dynamic Proteomics of Herpes Simplex Virus Infection
title_short	Dynamic Proteomics of Herpes Simplex Virus Infection
title_full	Dynamic Proteomics of Herpes Simplex Virus Infection
title_fullStr	Dynamic Proteomics of Herpes Simplex Virus Infection
title_full_unstemmed	Dynamic Proteomics of Herpes Simplex Virus Infection
title_sort	dynamic proteomics of herpes simplex virus infection
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/5a469c18db8f4c55bd316f119c67c6ab
work_keys_str_mv	AT nirdrayman dynamicproteomicsofherpessimplexvirusinfection AT omerkarin dynamicproteomicsofherpessimplexvirusinfection AT avimayo dynamicproteomicsofherpessimplexvirusinfection AT tamardanon dynamicproteomicsofherpessimplexvirusinfection AT levshapira dynamicproteomicsofherpessimplexvirusinfection AT dorrafael dynamicproteomicsofherpessimplexvirusinfection AT anatzimmer dynamicproteomicsofherpessimplexvirusinfection AT anatbren dynamicproteomicsofherpessimplexvirusinfection AT orenkobiler dynamicproteomicsofherpessimplexvirusinfection AT urialon dynamicproteomicsofherpessimplexvirusinfection
_version_	1718427276956139520

Dynamic Proteomics of Herpes Simplex Virus Infection

Ejemplares similares