A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry

Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Anh Phuong Luu, Zhenlan Yao, Sangeetha Ramachandran, Stephanie A. Azzopardi, Linde A. Miles, William M. Schneider, H.-Heinrich Hoffmann, Leonia Bozzacco, Gustavo Garcia, Danyang Gong, Robert Damoiseaux, Hengli Tang, Kouki Morizono, Charles M. Rudin, Ren Sun, Vaithilingaraja Arumugaswami, John T. Poirier, Margaret R. MacDonald, Charles M. Rice, Melody M. H. Li
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	CRISPR activation Zika virus proviral factors WWTR1 Rho GTPases RhoV Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2f3d203580654a47b1222b3bf06f5e6f
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2f3d203580654a47b1222b3bf06f5e6f
record_format	dspace
spelling	oai:doaj.org-article:2f3d203580654a47b1222b3bf06f5e6f2021-11-25T19:12:21ZA CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry10.3390/v131121131999-4915https://doaj.org/article/2f3d203580654a47b1222b3bf06f5e6f2021-10-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2113https://doaj.org/toc/1999-4915Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically targeted, we transcriptionally upregulated all known human coding genes with an engineered CRISPR–Cas9 activation complex in human fibroblasts deficient in interferon (IFN) signaling. We identified Ras homolog family member V (<i>RhoV</i>) and WW domain-containing transcription regulator 1 (<i>WWTR1</i>) as proviral factors, and found them to play important roles during early ZIKV infection in A549 cells. We then focused on RhoV, a Rho GTPase with atypical terminal sequences and membrane association, and validated its proviral effects on ZIKV infection and virion production in SNB-19 cells. We found that RhoV promotes infection of some flaviviruses and acts at the step of viral entry. Furthermore, RhoV proviral effects depend on the complete GTPase cycle. By depleting Rho GTPases and related proteins, we identified RhoB and Pak1 as additional proviral factors. Taken together, these results highlight the positive role of RhoV in ZIKV infection and confirm CRISPR activation as a relevant method to identify novel host–pathogen interactions.Anh Phuong LuuZhenlan YaoSangeetha RamachandranStephanie A. AzzopardiLinde A. MilesWilliam M. SchneiderH.-Heinrich HoffmannLeonia BozzaccoGustavo GarciaDanyang GongRobert DamoiseauxHengli TangKouki MorizonoCharles M. RudinRen SunVaithilingaraja ArumugaswamiJohn T. PoirierMargaret R. MacDonaldCharles M. RiceMelody M. H. LiMDPI AGarticleCRISPR activationZika virusproviral factorsWWTR1Rho GTPasesRhoVMicrobiologyQR1-502ENViruses, Vol 13, Iss 2113, p 2113 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR activation Zika virus proviral factors WWTR1 Rho GTPases RhoV Microbiology QR1-502
spellingShingle	CRISPR activation Zika virus proviral factors WWTR1 Rho GTPases RhoV Microbiology QR1-502 Anh Phuong Luu Zhenlan Yao Sangeetha Ramachandran Stephanie A. Azzopardi Linde A. Miles William M. Schneider H.-Heinrich Hoffmann Leonia Bozzacco Gustavo Garcia Danyang Gong Robert Damoiseaux Hengli Tang Kouki Morizono Charles M. Rudin Ren Sun Vaithilingaraja Arumugaswami John T. Poirier Margaret R. MacDonald Charles M. Rice Melody M. H. Li A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
description	Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically targeted, we transcriptionally upregulated all known human coding genes with an engineered CRISPR–Cas9 activation complex in human fibroblasts deficient in interferon (IFN) signaling. We identified Ras homolog family member V (<i>RhoV</i>) and WW domain-containing transcription regulator 1 (<i>WWTR1</i>) as proviral factors, and found them to play important roles during early ZIKV infection in A549 cells. We then focused on RhoV, a Rho GTPase with atypical terminal sequences and membrane association, and validated its proviral effects on ZIKV infection and virion production in SNB-19 cells. We found that RhoV promotes infection of some flaviviruses and acts at the step of viral entry. Furthermore, RhoV proviral effects depend on the complete GTPase cycle. By depleting Rho GTPases and related proteins, we identified RhoB and Pak1 as additional proviral factors. Taken together, these results highlight the positive role of RhoV in ZIKV infection and confirm CRISPR activation as a relevant method to identify novel host–pathogen interactions.
format	article
author	Anh Phuong Luu Zhenlan Yao Sangeetha Ramachandran Stephanie A. Azzopardi Linde A. Miles William M. Schneider H.-Heinrich Hoffmann Leonia Bozzacco Gustavo Garcia Danyang Gong Robert Damoiseaux Hengli Tang Kouki Morizono Charles M. Rudin Ren Sun Vaithilingaraja Arumugaswami John T. Poirier Margaret R. MacDonald Charles M. Rice Melody M. H. Li
author_facet	Anh Phuong Luu Zhenlan Yao Sangeetha Ramachandran Stephanie A. Azzopardi Linde A. Miles William M. Schneider H.-Heinrich Hoffmann Leonia Bozzacco Gustavo Garcia Danyang Gong Robert Damoiseaux Hengli Tang Kouki Morizono Charles M. Rudin Ren Sun Vaithilingaraja Arumugaswami John T. Poirier Margaret R. MacDonald Charles M. Rice Melody M. H. Li
author_sort	Anh Phuong Luu
title	A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
title_short	A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
title_full	A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
title_fullStr	A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
title_full_unstemmed	A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry
title_sort	crispr activation screen identifies an atypical rho gtpase that enhances zika viral entry
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/2f3d203580654a47b1222b3bf06f5e6f
work_keys_str_mv	AT anhphuongluu acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT zhenlanyao acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT sangeetharamachandran acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT stephanieaazzopardi acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT lindeamiles acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT williammschneider acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT hheinrichhoffmann acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT leoniabozzacco acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT gustavogarcia acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT danyanggong acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT robertdamoiseaux acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT henglitang acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT koukimorizono acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT charlesmrudin acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT rensun acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT vaithilingarajaarumugaswami acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT johntpoirier acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT margaretrmacdonald acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT charlesmrice acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT melodymhli acrispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT anhphuongluu crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT zhenlanyao crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT sangeetharamachandran crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT stephanieaazzopardi crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT lindeamiles crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT williammschneider crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT hheinrichhoffmann crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT leoniabozzacco crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT gustavogarcia crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT danyanggong crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT robertdamoiseaux crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT henglitang crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT koukimorizono crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT charlesmrudin crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT rensun crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT vaithilingarajaarumugaswami crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT johntpoirier crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT margaretrmacdonald crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT charlesmrice crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry AT melodymhli crispractivationscreenidentifiesanatypicalrhogtpasethatenhanceszikaviralentry
_version_	1718410144613662720

A CRISPR Activation Screen Identifies an Atypical Rho GTPase That Enhances Zika Viral Entry

Ejemplares similares