A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response

ABSTRACT The type I interferon (IFN) signaling pathway restricts infection of many divergent families of RNA and DNA viruses by inducing hundreds of IFN-stimulated genes (ISGs), some of which have direct antiviral activity. We screened 813 short hairpin RNA (shRNA) constructs targeting 245 human ISG...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jianqing Li, Steve C. Ding, Hyelim Cho, Brian C. Chung, Michael Gale, Sumit K. Chanda, Michael S. Diamond
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/1f4c3ee4bf064ce890295a72819b2487
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:1f4c3ee4bf064ce890295a72819b2487
record_format	dspace
spelling	oai:doaj.org-article:1f4c3ee4bf064ce890295a72819b24872021-11-15T15:40:05ZA Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response10.1128/mBio.00385-132150-7511https://doaj.org/article/1f4c3ee4bf064ce890295a72819b24872013-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00385-13https://doaj.org/toc/2150-7511ABSTRACT The type I interferon (IFN) signaling pathway restricts infection of many divergent families of RNA and DNA viruses by inducing hundreds of IFN-stimulated genes (ISGs), some of which have direct antiviral activity. We screened 813 short hairpin RNA (shRNA) constructs targeting 245 human ISGs using a flow cytometry approach to identify genes that modulated infection of West Nile virus (WNV) in IFN-β-treated human cells. Thirty ISGs with inhibitory effects against WNV were identified, including several novel genes that had antiviral activity against related and unrelated positive-strand RNA viruses. We also defined one ISG, activating signal cointegrator complex 3 (ASCC3), which functioned as a negative regulator of the host defense response. Silencing of ASCC3 resulted in upregulation of multiple antiviral ISGs, which correlated with inhibition of infection of several positive-strand RNA viruses. Reciprocally, ectopic expression of human ASCC3 or mouse Ascc3 resulted in downregulation of ISGs and increased viral infection. Mechanism-of-action and RNA sequencing studies revealed that ASCC3 functions to modulate ISG expression in an IRF-3- and IRF-7-dependent manner. Compared to prior ectopic ISG expression studies, our shRNA screen identified novel ISGs that restrict infection of WNV and other viruses and defined a new counterregulatory ISG, ASCC3, which tempers cell-intrinsic immunity. IMPORTANCE West Nile virus (WNV) is a mosquito-transmitted virus that continues to pose a threat to public health. Innate immune responses, especially those downstream of type I interferon (IFN) signaling, are critical for controlling virus infection and spread. We performed a genetic screen using a gene silencing approach and identified 30 interferon-stimulated genes (ISGs) that contributed to the host antiviral response against WNV. As part of this screen, we also identified a novel negative regulatory protein, ASCC3, which dampens expression of ISGs, including those with antiviral or proinflammatory activity. In summary, our studies define a series of heretofore-uncharacterized ISGs with antiviral effects against multiple viruses or counterregulatory effects that temper IFN signaling and likely minimize immune-mediated pathology.Jianqing LiSteve C. DingHyelim ChoBrian C. ChungMichael GaleSumit K. ChandaMichael S. DiamondAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 3 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Jianqing Li Steve C. Ding Hyelim Cho Brian C. Chung Michael Gale Sumit K. Chanda Michael S. Diamond A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
description	ABSTRACT The type I interferon (IFN) signaling pathway restricts infection of many divergent families of RNA and DNA viruses by inducing hundreds of IFN-stimulated genes (ISGs), some of which have direct antiviral activity. We screened 813 short hairpin RNA (shRNA) constructs targeting 245 human ISGs using a flow cytometry approach to identify genes that modulated infection of West Nile virus (WNV) in IFN-β-treated human cells. Thirty ISGs with inhibitory effects against WNV were identified, including several novel genes that had antiviral activity against related and unrelated positive-strand RNA viruses. We also defined one ISG, activating signal cointegrator complex 3 (ASCC3), which functioned as a negative regulator of the host defense response. Silencing of ASCC3 resulted in upregulation of multiple antiviral ISGs, which correlated with inhibition of infection of several positive-strand RNA viruses. Reciprocally, ectopic expression of human ASCC3 or mouse Ascc3 resulted in downregulation of ISGs and increased viral infection. Mechanism-of-action and RNA sequencing studies revealed that ASCC3 functions to modulate ISG expression in an IRF-3- and IRF-7-dependent manner. Compared to prior ectopic ISG expression studies, our shRNA screen identified novel ISGs that restrict infection of WNV and other viruses and defined a new counterregulatory ISG, ASCC3, which tempers cell-intrinsic immunity. IMPORTANCE West Nile virus (WNV) is a mosquito-transmitted virus that continues to pose a threat to public health. Innate immune responses, especially those downstream of type I interferon (IFN) signaling, are critical for controlling virus infection and spread. We performed a genetic screen using a gene silencing approach and identified 30 interferon-stimulated genes (ISGs) that contributed to the host antiviral response against WNV. As part of this screen, we also identified a novel negative regulatory protein, ASCC3, which dampens expression of ISGs, including those with antiviral or proinflammatory activity. In summary, our studies define a series of heretofore-uncharacterized ISGs with antiviral effects against multiple viruses or counterregulatory effects that temper IFN signaling and likely minimize immune-mediated pathology.
format	article
author	Jianqing Li Steve C. Ding Hyelim Cho Brian C. Chung Michael Gale Sumit K. Chanda Michael S. Diamond
author_facet	Jianqing Li Steve C. Ding Hyelim Cho Brian C. Chung Michael Gale Sumit K. Chanda Michael S. Diamond
author_sort	Jianqing Li
title	A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
title_short	A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
title_full	A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
title_fullStr	A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
title_full_unstemmed	A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response
title_sort	short hairpin rna screen of interferon-stimulated genes identifies a novel negative regulator of the cellular antiviral response
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/1f4c3ee4bf064ce890295a72819b2487
work_keys_str_mv	AT jianqingli ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT stevecding ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT hyelimcho ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT briancchung ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT michaelgale ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT sumitkchanda ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT michaelsdiamond ashorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT jianqingli shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT stevecding shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT hyelimcho shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT briancchung shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT michaelgale shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT sumitkchanda shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse AT michaelsdiamond shorthairpinrnascreenofinterferonstimulatedgenesidentifiesanovelnegativeregulatorofthecellularantiviralresponse
_version_	1718427751338213376

A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral Response

Ejemplares similares