# Characterizing host proteins that restrict Zika virus

> **NIH NIH F30** · UNIVERSITY OF WASHINGTON · 2024 · $49,802

## Abstract

PROJECT SUMMARY / ABSTRACT
 Zika virus (ZIKV) re-emerged over the last several decades to cause large outbreaks in the Asian Pacific
and Americas. ZIKV disease, though usually mild, can occasionally cause severe neurological complications,
including developmental defects in babies born to a ZIKV-infected person. Much remains to be learned about
ZIKV host-pathogen interactions, including how the innate immune system fights the early stages of infection in
human cells. Given their role in inhibiting viral replication, innate immune factors could help explain the
heterogeneity in ZIKV clinical outcome and could lead to the development of treatments for ZIKV. The type I
interferon (IFN) response plays a particularly critical role in the innate immune response to viruses, including
ZIKV. IFN is secreted from virus-infected cells, binds its receptor on nearby cells, and sets off a signaling pathway
that culminates in a transcriptional program turning on hundreds of interferon-stimulated genes (ISGs), some of
which encode antiviral proteins against a given virus. While the IFN response is clearly important in restricting
ZIKV, systematic studies to define which host genes are responsible for this potent effect are lacking. While
several specific antiviral ISGs have been identified, there have not been broader attempts to define all host
genes that contribute to IFN restriction of ZIKV, including non-ISGs that may play a regulatory role in the pathway.
Our lab performed a CRISPR knockout screen to identify genes that contribute to IFN restriction of ZIKV. The
screen approach successfully identified genes involved in IFN signaling (positive controls). IFI6, a previously
described ISG and flavivirus restriction factor, was also identified, as was AMOTL2, a non-ISG in our cell type
and a gene with no described role in innate immunity. Despite the fact that AMOTL2 is not itself IFN-induced,
AMOTL2 knockout increased ZIKV replication in the presence, but not absence, of IFN. These findings confirmed
the IFN-specific antiviral phenotype of AMOTL2 and led to the hypothesis that AMOTL2 acts upstream of ISG
transcription in the IFN response. My preliminary results support this model and are the basis for the proposed
studies. In Aim 1, the mechanism of AMOTL2’s viral restriction will be better elucidated. This will be achieved by
first validating the specificity of the observed phenotype through rescue experiments, then assessing the
involvement of AMOTL2’s binding partners YAP and TAZ in IFN restriction of ZIKV through double knockouts
and co-IP assays. In Aim 2, I will test the breadth of AMOTL2’s antiviral phenotype across diverse viruses (Aim
2a) and with ZIKV infections in a biologically-relevant primary cell type (Aim 2b). The complementary results of
my two aims will enhance our understanding of the IFN response against ZIKV and possibly other viruses.

## Key facts

- **NIH application ID:** 10995008
- **Project number:** 1F30AI181359-01A1
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Alexandra Claiborne Willcox
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $49,802
- **Award type:** 1
- **Project period:** 2024-08-01 → 2026-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10995008

## Citation

> US National Institutes of Health, RePORTER application 10995008, Characterizing host proteins that restrict Zika virus (1F30AI181359-01A1). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10995008. Licensed CC0.

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