Genome-wide RNAi screen reveals a specific sensitivity of IRES-containing RNA viruses to host translation inhibition

Cherry, Sara; Doukas, Tammy; Armknecht, Susan; Whelan, Sean P. J.; Wang, Hui; Sarnow, Peter; Perrimon, Norbert

doi:10.1101/gad.1267905

Cited by 199 publications

(182 citation statements)

References 25 publications

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“…org, and RNAi was performed as described (17). The experimental methods and image analysis have been extensively described (26,27,53). In brief, cells were passaged into serum-free media (SFM) and plated into wells containing dsRNA at 250 ng/16,500 cells in 384-well plates.…”

Section: Methodsmentioning

confidence: 99%

“…For the U0126 experiments, food and virus was changed every 3 d. For the insulin experiments, insulin was only provided for the first 3 d, subsequently food and virus was changed every 3 d. For immunoblots, 15 fly guts were dissected in PBS and processed in RIPA buffer, protease inhibitor mixture (PP2, PP3) and PMSF as described (17). For RNA, 15 fly guts were dissected in PBS and then processed by using TRIzol as per the manufacturer's protocol and as described (53). For microscopy, fly guts were dissected as described (55).…”

Section: Methodsmentioning

confidence: 99%

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ERK signaling couples nutrient status to antiviral defense in the insect gut

Hopkins

Sabin

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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127

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A unique facet of arthropod-borne virus (arbovirus) infection is that the pathogens are orally acquired by an insect vector during the taking of a blood meal, which directly links nutrient acquisition and pathogen challenge. We show that the nutrient responsive ERK pathway is both induced by and restricts disparate arboviruses in Drosophila intestines, providing insight into the molecular determinants of the antiviral "midgut barrier." Wild-type flies are refractory to oral infection by arboviruses, including Sindbis virus and vesicular stomatitis virus, but this innate restriction can be overcome chemically by oral administration of an ERK pathway inhibitor or genetically via the specific loss of ERK in Drosophila intestinal epithelial cells. In addition, we found that vertebrate insulin, which activates ERK in the mosquito gut during a blood meal, restricts viral infection in Drosophila cells and against viral invasion of the insect gut epithelium. We find that ERK's antiviral signaling activity is likely conserved in Aedes mosquitoes, because genetic or pharmacologic manipulation of the ERK pathway affects viral infection of mosquito cells. These studies demonstrate that ERK signaling has a broadly antiviral role in insects and suggest that insects take advantage of cross-species signals in the meal to trigger antiviral immunity.innate immunity | enterocytes

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

ERK signaling couples nutrient status to antiviral defense in the insect gut

Hopkins

Sabin

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

127

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“…The advent of siRNA technology and the availability of genome-wide siRNA libraries have been useful in identifying host factors required for influenza virus, an NS RNA virus, and several positive-strand RNA viruses, as well as HIV (10)(11)(12)(13)(14)(15)(16)(17)(18)(19). The lack of similar studies with other NS RNA viruses has limited the understanding of the role of host cell factors in replication of these viruses.…”

mentioning

confidence: 99%

RNAi screening reveals requirement for host cell secretory pathway in infection by diverse families of negative-strand RNA viruses

Panda

Das

Dinh

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Negative-strand (NS) RNA viruses comprise many pathogens that cause serious diseases in humans and animals. Despite their clinical importance, little is known about the host factors required for their infection. Using vesicular stomatitis virus (VSV), a prototypic NS RNA virus in the family Rhabdoviridae, we conducted a human genomewide siRNA screen and identified 72 host genes required for viral infection. Many of these identified genes were also required for infection by two other NS RNA viruses, the lymphocytic choriomeningitis virus of the Arenaviridae family and human parainfluenza virus type 3 of the Paramyxoviridae family. Genes affecting different stages of VSV infection, such as entry/uncoating, gene expression, and assembly/release, were identified. Depletion of the proteins of the coatomer complex I or its upstream effectors ARF1 or GBF1 led to detection of reduced levels of VSV RNA. Coatomer complex I was also required for infection of lymphocytic choriomeningitis virus and human parainfluenza virus type 3. These results highlight the evolutionarily conserved requirements for gene expression of diverse families of NS RNA viruses and demonstrate the involvement of host cell secretory pathway in the process.RNA interference | transcription and replication | host cell factors for virus infection

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“…In the past, classical virology focused on the virus itself, ignoring cellular functions and complex biological processes in the cell as an RNA viruses. This study suggests another direction for antiviral drug discovery by modulating the host cell's ribosome activity [57].…”

Section: Rnai Screens To Identify Host Factors That Influence Viral Imentioning

confidence: 83%

Illuminating the host – How RNAi screens shed light on host‐pathogen interactions

Prudêncio

Lehmann

2009

Biotechnology Journal

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Over millions of years pathogens have coevolved with their respective hosts utilizing host cell functions for survival and replication. Despite remarkable progress in developing antibiotics and vaccination strategies in the last century, infectious diseases still remain a severe threat to human health. Meanwhile, genomic research offers a new era of data‐generating platforms that will dramatically enhance our knowledge of pathogens and the diseases they cause. Improvements in gene knockdown studies by RNA interference (RNAi) combined with recent developments in instrumentation and image analysis enable the use of high‐throughput screening approaches to elucidate host gene functions exploited by pathogens. Although only a few RNAi‐based screens focusing on host genes have been reported so far, these studies have already uncovered hundreds of genes not previously known to be involved in pathogen infection. This review describes recent progress in RNAi screening approaches, highlighting both the limitations and the tremendous potential of RNAi‐based screens for the identification of essential host cell factors during infection.

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Genome-wide RNAi screen reveals a specific sensitivity of IRES-containing RNA viruses to host translation inhibition

Cited by 199 publications

References 25 publications

ERK signaling couples nutrient status to antiviral defense in the insect gut

ERK signaling couples nutrient status to antiviral defense in the insect gut

RNAi screening reveals requirement for host cell secretory pathway in infection by diverse families of negative-strand RNA viruses

Illuminating the host – How RNAi screens shed light on host‐pathogen interactions

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