# Elucidating the mechanisms of viral life cycles under near-native conditions

> **NIH NIH DP2** · VANDERBILT UNIVERSITY · 2021 · $1,426,500

## Abstract

Project Summary
Mononegavirales is a taxonomic order of viruses, so classified for their negative sense single
stranded RNA genome and their pleomorphic membrane-enveloped virions. Mononegavirus life
cycles involve a number of different events, including entry of virions into host cells, viral mRNA
transcription, genome replication and virus assembly, and viral budding from host cells. These
events are carried out through molecular interactions between virus and host cell machinery;
elucidating these interactions is key to understanding viral life cycles and identifying potential
therapeutic targets.
Studies of viral machinery are typically limited to isolated particles or assemblies; this removes
them from their native environments and strips away important molecular interactions. To
preserve biological context, viral machinery must be studied in situ, i.e. under near-native
conditions, such as within intact virions or cells. These environments are a complex, disordered
mixture of molecules, making it particularly difficult to obtain molecular resolution information.
Here, we propose to study three mononegaviruses: measles, rabies, and Ebola viruses. Each
serve as prototypical viruses for their taxonomic families, and each are pathogens important to
global health.
To carry out our proposed research, we will use and develop in situ structural biology methods.
Our primary method will be cryo-electron tomography (cryo-ET), a type of cryo-electron
microscopy (cryo-EM) that allows for visualization of three-dimensional volumes. This
overcomes the typical cryo-EM requirement of thin monolayers of purified particles, allowing for
the acquisition of molecular-resolution information in near-native environments. We will develop
data collection and computational methods for cryo-ET to enable rapid, automated data
collection, high-resolution structure determination, and accurate molecular identification. We will
also use and develop methods complementary to cryo-ET including focused ion-beam milling
and correlative light and electron microscopy approaches.
Our research will provide novel biological insights three important viruses, but more broadly, it
will demonstrate a transformative approach for studying viruses. Rather than trying to tease
apart function and interactions through indirect biochemical means, our research will provide an
infrastructure to directly observe the virus and host cell machinery with complete biological
contexts under near-native conditions.

## Key facts

- **NIH application ID:** 10242476
- **Project number:** 1DP2GM146321-01
- **Recipient organization:** VANDERBILT UNIVERSITY
- **Principal Investigator:** William N Wan
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $1,426,500
- **Award type:** 1
- **Project period:** 2021-09-24 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10242476

## Citation

> US National Institutes of Health, RePORTER application 10242476, Elucidating the mechanisms of viral life cycles under near-native conditions (1DP2GM146321-01). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10242476. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*