# Cell Biology of Reovirus Infection

> **NIH NIH R01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2021 · $518,440

## Abstract

Most viruses that replicate in the cytoplasm of host cells form neoorganelles that serve as sites of viral
genome replication and particle assembly. These highly specialized inclusion structures concentrate viral
replication proteins and nucleic acids, prevent activation of cell-intrinsic defenses, and coordinate release of
progeny particles. Despite the importance of inclusion complexes in viral replication, there are key gaps in
knowledge about how these organelles form and mediate their functions. The proposed research uses reovirus,
a genetically tractable virus that has been linked to celiac disease and shows promise for oncolytic applications,
to elucidate mechanisms of double-stranded (ds) RNA virus inclusion formation, genome replication, and
progeny particle assembly and release. Like other dsRNA viruses, which include important pathogens of
animals (orbiviruses) and humans (rotaviruses), reovirus inclusions are nucleated by viral nonstructural proteins
that recruit viral structural proteins for genome replication and particle assembly. We have discovered that (i)
reovirus inclusions originate from the endoplasmic reticulum (ER), (ii) capsid assembly requires the TRiC
chaperonin, and (iii) progeny particles are transported from inclusions and released using a vesicular sorting
mechanism dependent on modified lysosomes and actin. Three integrated specific aims are proposed to fill key
knowledge gaps about reovirus inclusion biogenesis, capsid assembly, and particle egress. In Specific Aim 1,
functions of reovirus nonstructural proteins σNS and µNS in inclusion formation and genome synthesis will be
elucidated using biochemical and cell-imaging approaches. The structure of σNS will be determined to facilitate
analysis of its function in ER reorganization and dsRNA synthesis. In Specific Aim 2, interactions between
TRiC and reovirus outer-capsid protein σ3 will be defined using mass spectrometry and cryo-EM analysis. The
function of the TRiC-loading protein, prefoldin, in σ3 maturation will be defined using in vitro protein-folding
assays. The protein-folding network required to assemble σ3 onto newly formed virions will be defined using
gene-targeting and biochemical assays. In Specific Aim 3, the lipid-biosynthetic and transport pathways
required for reovirus egress will be elucidated by defining a function for Niemann-Pick disease type C1 protein,
identified in RNA interference and CRISPR screens, in the egress process. The mechanism by which mature
virions are transferred into membranous, multilamellar structures and then to smaller membranous carriers to
reach the plasma membrane will be defined using light and electron microscopy. The function of actin and
myosin 9 in inclusion biogenesis, particle assembly, and viral egress will be defined using pharmacologic
inhibitors and cell imaging. These studies will enhance an understanding of mechanisms by which pathogenic
viruses alter cellular architecture to engineer inclusion organelles, ...

## Key facts

- **NIH application ID:** 10132965
- **Project number:** 5R01AI032539-30
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** TERENCE S. DERMODY
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $518,440
- **Award type:** 5
- **Project period:** 1992-07-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10132965

## Citation

> US National Institutes of Health, RePORTER application 10132965, Cell Biology of Reovirus Infection (5R01AI032539-30). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10132965. Licensed CC0.

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