A Systematic Analysis of Host Factors Reveals a Med23-Interferon-λ Regulatory Axis against Herpes Simplex Virus Type 1 Replication

PLoS Pathogens

Jürgen Haas¹  Rui Chen²  Kim Martin²  Marie H. Craigon²  Lakshmi N. Kaza²  Even Fossum²  Peter Ghazal²  Samantha J. Griffiths²  Helen L. Zenner³  Stacey Efstathiou³  Colin M. Crump³  Ralf Zimmer⁴  Orland Gonzalez⁴  Caroline C. Friedel⁴  Chris Boutell⁵  Manfred Koegl⁶  Gerald Barry⁷  John K. Fazakerley⁷  Antonio Volpi⁸  Francesca Pica⁸ 

[1] Centre for Systems Biology at Edinburgh, University of Edinburgh, Edinburgh, United Kingdom;Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom;Division of Virology, Department of Pathology Cambridge University, Cambridge, United Kingdom;Institute for Informatics, Ludwig-Maximilians Universität München, München, Germany;MRC-University of Glasgow Centre for Virus Research, Glasgow, United Kingdom;Preclinical Target Development and Genomics and Proteomics Core Facilities, German Cancer Research Center, Heidelberg, Germany;The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, United Kingdom;University of Rome Tor Vergata, Rome, Italy

关键词: Small interfering RNAs;    Viral replication;    Protein interactions;    Interferons;    Transfection;    HeLa cells;    RNA interference;    Transcription factors;   

DOI  :  10.1371/journal.ppat.1003514

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Herpes simplex virus type 1 (HSV-1) is a neurotropic virus causing vesicular oral or genital skin lesions, meningitis and other diseases particularly harmful in immunocompromised individuals. To comprehensively investigate the complex interaction between HSV-1 and its host we combined two genome-scale screens for host factors (HFs) involved in virus replication. A yeast two-hybrid screen for protein interactions and a RNA interference (RNAi) screen with a druggable genome small interfering RNA (siRNA) library confirmed existing and identified novel HFs which functionally influence HSV-1 infection. Bioinformatic analyses found the 358 HFs were enriched for several pathways and multi-protein complexes. Of particular interest was the identification of Med23 as a strongly anti-viral component of the largely pro-viral Mediator complex, which links specific transcription factors to RNA polymerase II. The anti-viral effect of Med23 on HSV-1 replication was confirmed in gain-of-function gene overexpression experiments, and this inhibitory effect was specific to HSV-1, as a range of other viruses including Vaccinia virus and Semliki Forest virus were unaffected by Med23 depletion. We found Med23 significantly upregulated expression of the type III interferon family (IFN-λ) at the mRNA and protein level by directly interacting with the transcription factor IRF7. The synergistic effect of Med23 and IRF7 on IFN-λ induction suggests this is the major transcription factor for IFN-λ expression. Genotypic analysis of patients suffering recurrent orofacial HSV-1 outbreaks, previously shown to be deficient in IFN-λ secretion, found a significant correlation with a single nucleotide polymorphism in the IFN-λ3 (IL28b) promoter strongly linked to Hepatitis C disease and treatment outcome. This paper describes a link between Med23 and IFN-λ, provides evidence for the crucial role of IFN-λ in HSV-1 immune control, and highlights the power of integrative genome-scale approaches to identify HFs critical for disease progression and outcome.

【 授权许可】

CC BY