Haploid Screening for the Identification of Host Factors in Virus Infection

ABSTRACTThroughout the rabies virus (RABV) infectious cycle, host-virus interactions define its capacity to replicate, escape the immune response, and spread. As phosphorylation is a key regulatory mechanism involved in most cellular processes, kinases represent a target of choice to identify host factors required for viral replication. A kinase and phosphatase small interfering RNA (siRNA) high-content screening was performed on a fluorescent protein-recombinant field isolate (Tha RABV). We identified 57 high-confidence key host factors important for RABV replication with a readout set at 18 h postinfection and 73 with a readout set at 36 h postinfection, including 24 common factors at all stages of the infection. Amongst them, gene clusters of the most prominent pathways were determined. Up to 15 mitogen-activated protein kinases (MAPKs) and effectors, including MKK7 (associated with Jun N-terminal protein kinase [JNK] signalization) and DUSP5, as well as 17 phosphatidylinositol (PI)-related proteins, including PIP5K1C and MTM1, were found to be involved in the later stage of RABV infection. The importance of these pathways was further validated, as small molecules Ro 31-8820 and PD 198306 inhibited RABV replication in human neurons.IMPORTANCERabies virus relies on cellular machinery for its replication while simultaneously evading the host immune response. Despite their importance, little is known about the key host factors required for rabies virus infection. Here, we focused on the human kinome, at the core of many cellular pathways, to unveil a new understanding of the rabies virus infectious cycle and to discover new potential therapeutic targets in a small interfering RNA screening. The mitogen-activated protein kinase pathway and phosphatidylinositol metabolism were identified as prominent factors involved in rabies virus infection, and those findings were further confirmed in human neurons. While bringing a new insight into rabies virus biology, we also provide a new list of host factors involved in rabies virus infection.

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A Genome-Wide Small Interfering RNA Screen Identifies Host Factors Required for Vesicular Stomatitis Virus Infection

Journal of Virology ◽

10.1128/jvi.00642-14 ◽

2014 ◽

Vol 88 (15) ◽

pp. 8355-8360 ◽

Cited By ~ 19

Author(s):

A. S.-Y. Lee ◽

R. Burdeinick-Kerr ◽

S. P. J. Whelan

Keyword(s):

Virus Infection ◽

Vesicular Stomatitis Virus ◽

Small Interfering Rna ◽

Host Factors ◽

Genome Wide ◽

A Genome ◽

Vesicular Stomatitis ◽

Interfering Rna

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Host factors associated with the kinetics of Epstein-Barr virus DNA load in patients with primary Epstein-Barr virus infection

Microbiology and Immunology ◽

10.1111/j.1348-0421.2011.00410.x ◽

2012 ◽

Vol 56 (2) ◽

pp. 93-98 ◽

Cited By ~ 3

Author(s):

Hidetaka Nakai ◽

Yoshiki Kawamura ◽

Ken Sugata ◽

Hiroko Sugiyama ◽

Yoshihiko Enomoto ◽

...

Keyword(s):

Virus Infection ◽

Epstein Barr Virus ◽

Host Factors ◽

Epstein Barr Virus Infection ◽

Factors Associated ◽

Barr Virus ◽

Epstein Barr ◽

Barr Virus Infection ◽

Kinetics Of

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The German Hep-Net Acute Hepatitis C Cohort: Impact of Viral and Host Factors on the Initial Presentation of Acute Hepatitis C Virus Infection

Zeitschrift für Gastroenterologie ◽

10.1055/s-0028-1109149 ◽

2009 ◽

Vol 47 (06) ◽

pp. 531-540 ◽

Cited By ~ 22

Author(s):

K. Deterding ◽

J. Wiegand ◽

N. Grüner ◽

A. Hahn ◽

E. Jäckel ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Virus Infection ◽

Acute Hepatitis ◽

Initial Presentation ◽

Host Factors ◽

Acute Hepatitis C ◽

Hepatitis C Virus Infection

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Impact of cohort size and host factors on combined analysis of interleukin 28B rs12979860 and rs8099917 in hepatitis C virus infection

Hepatology ◽

10.1002/hep.25923 ◽

2013 ◽

Vol 57 (1) ◽

pp. 416-417 ◽

Cited By ~ 2

Author(s):

Janett Fischer ◽

Stephan Böhm ◽

Jacob George ◽

Christoph Sarrazin ◽

Thomas Berg

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Virus Infection ◽

Host Factors ◽

Cohort Size ◽

Hepatitis C Virus Infection ◽

Combined Analysis ◽

Interleukin 28B

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A Comprehensive View on the Host Factors and Viral Proteins Associated With Porcine Epidemic Diarrhea Virus Infection

Frontiers in Microbiology ◽

10.3389/fmicb.2021.762358 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yi Hu ◽

Xiaohong Xie ◽

Lingchen Yang ◽

Aibing Wang

Keyword(s):

Virus Infection ◽

Porcine Epidemic Diarrhea Virus ◽

Molecular Mechanisms ◽

Viral Proteins ◽

Host Factors ◽

Watery Diarrhea ◽

Economic Losses ◽

Target Cells ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV), a coronavirus pathogen of the pig intestinal tract, can cause fatal watery diarrhea in piglets, thereby causing huge economic losses to swine industries around the world. The pathogenesis of PEDV has intensively been studied; however, the viral proteins of PEDV and the host factors in target cells, as well as their interactions, which are the foundation of the molecular mechanisms of viral infection, remain to be summarized and updated. PEDV has multiple important structural and functional proteins, which play various roles in the process of virus infection. Among them, the S and N proteins play vital roles in biological processes related to PEDV survival via interacting with the host cell proteins. Meanwhile, a number of host factors including receptors are required for the infection of PEDV via interacting with the viral proteins, thereby affecting the reproduction of PEDV and contributing to its life cycle. In this review, we provide an updated understanding of viral proteins and host factors, as well as their interactions in terms of PEDV infection. Additionally, the effects of cellular factors, events, and signaling pathways on PEDV infection are also discussed. Thus, these comprehensive and profound insights should facilitate for the further investigations, control, and prevention of PEDV infection.

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The Lipid Transfer Protein 1 from Nicotiana benthamiana Assists Bamboo mosaic virus Accumulation

Viruses ◽

10.3390/v12121361 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1361

Author(s):

Ling-Ying Chiu ◽

I-Hsuan Chen ◽

Yau-Heiu Hsu ◽

Ching-Hsiu Tsai

Keyword(s):

Virus Infection ◽

Mosaic Virus ◽

Nicotiana Benthamiana ◽

Lipid Transfer Protein ◽

Lipid Binding ◽

Host Factors ◽

Functional Domain ◽

Lipid Transfer ◽

Calmodulin Binding ◽

Transfer Protein

Host factors play a pivotal role in regulating virus infection. Uncovering the mechanism of how host factors are involved in virus infection could pave the way to defeat viral disease. In this study, we characterized a lipid transfer protein, designated NbLTP1 in Nicotiana benthamiana, which was downregulated after Bamboo mosaic virus (BaMV) inoculation. BaMV accumulation significantly decreased in NbLTP1-knockdown leaves and protoplasts compared with the controls. The subcellular localization of the NbLTP1-orange fluorescent protein (OFP) was mainly the extracellular matrix. However, when we removed the signal peptide (NbLTP1/ΔSP-OFP), most of the expressed protein targeted chloroplasts. Both NbLTP1-OFP and NbLTP1/ΔSP-OFP were localized in chloroplasts when we removed the cell wall. These results suggest that NbLTP1 may have a secondary targeting signal. Transient overexpression of NbLTP1 had no effect on BaMV accumulation, but that of NbLTP1/ΔSP significantly increased BaMV expression. NbLTP1 may be a positive regulator of BaMV accumulation especially when its expression is associated with chloroplasts, where BaMV replicates. The mutation was introduced to the predicted phosphorylation site to simulate the phosphorylated status, NbLTP/ΔSP/P(+), which could still assist BaMV accumulation. By contrast, a mutant lacking calmodulin-binding or simulates the phosphorylation-negative status could not support BaMV accumulation. The lipid-binding activity of LTP1 was reported to be associated with calmodulin-binding and phosphorylation, by which the C-terminus functional domain of NbLTP1 may play a critical role in BaMV accumulation.

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