# DEFINING NOVEL, EVOLUTIONARILY CONSERVED HOST FACTORS CRITICAL FOR VIRUS INFECTION.

> **NIH NIH R01** · WASHINGTON UNIVERSITY · 2021 · $393,750

## Abstract

SUMMARY
 The goal of this project is to identify and characterize evolutionarily conserved host genes that are
important for virus infection in organisms from humans to nematodes by exploiting a unique C. elegans virus
infection system. C. elegans has played a key role in many fundamental discoveries in biology, but its use in
the study of viral pathogenesis has been limited by the lack of known viruses capable of naturally infecting C.
elegans. The recent discovery of Orsay Virus (ORV), the first and to date only known virus of C. elegans,
provide a novel route for genetic dissection of host factors essential for virus infection. In preliminary data, a
chemical mutagenesis screen in C. elegans was developed that identified two host genes essential for ORV
infection. The first gene, sid-3, encodes a non-receptor tyrosine kinase that is orthologous to the human
“activated Cdc42 associated kinase” (ACK1/TNK2). The second gene, viro-2, encodes an ortholog of human
Wiskott Aldrich syndrome proteins (N-WASP). ORV is a non-enveloped positive sense RNA virus; among viral
families with human pathogens, ORV is most closely related to the Picorna-, Calici- and Astroviridae. Deletion
of either TNK2 or N-WASP in human A549 cells led to ~1000-fold reduced titers of encephalomycarditis virus
(EMCV). ~100-fold reduction in coxsackie virus B3 (CVB3) was also observed in TNK2 deleted cells. Strikingly,
N-WASP is a known substrate for the kinase activity of TNK2, suggesting that these genes may comprise a
key pathway utilized by viruses. In support of this possibility, a third gene in C. elegans, nck-1, which is the
ortholog of the human NCK protein that binds to both TNK2 and N-WASP, was also found to be essential for
ORV infection. TNK2 has not been previously implicated in virus infection and represents a novel mammalian
pro-viral host factor discovered by genetic screening in C. elegans. The aims are: 1) Utilize C. elegans as a
discovery engine to comprehensively identify and characterize host factors critical for ORV infection. 2) To gain
mechanistic insight into TNK2 and N-WASP, the stage of EMCV and CVB3 infection impacted by these genes
and their essential protein domains and substrates required for their pro-viral functions will be defined. In
addition, among the C. elegans candidate genes identified in Aim 1 that have human orthologs, a subset that
phenocopy sid-3 and viro-2, which are the most likely candidates to act in the same pathway, will be prioritized
for similar analyses. 3) Finally, the effect of TNK2 deletion in mice on EMCV and CVB3 infection will be
defined. This proposal will yield novel insights into fundamental host genes and pathways necessary for virus
infection which could be novel targets for antiviral development.

## Key facts

- **NIH application ID:** 10141190
- **Project number:** 5R01AI134967-04
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** DAVID WANG
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $393,750
- **Award type:** 5
- **Project period:** 2018-05-02 → 2023-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10141190

## Citation

> US National Institutes of Health, RePORTER application 10141190, DEFINING NOVEL, EVOLUTIONARILY CONSERVED HOST FACTORS CRITICAL FOR VIRUS INFECTION. (5R01AI134967-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10141190. Licensed CC0.

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