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Principal Investigator: Christine Anne Roden
Organization: DUKE UNIVERSITY
Fiscal Year: 2024
Award: $38,033
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract:
INTRODUCTION: RNA viruses such as SARS-CoV-2 represent an ongoing public health threat. To replicate
properly, RNA viruses must perform multiple functions in the same cytoplasm (i.e., genome replication,
circularization, generation of sub-genomic RNA, packaging etc.) and for betacoronaviruses like SARS-CoV-2
the structural nucleocapsid protein (N) is required for all of these processes. How are RNA viruses able to
perform multiple functions in the same cytoplasm with their limited proteome (for example ~24 proteins for
SARS-CoV-2)? I hypothesize that one way RNA viruses achieve biochemical complexity is by condensation of
RNA and protein components. Biochemical complexity can emerge by virtue of the co-condensing RNA as
different viral RNA sequences confer different condensate properties. In this way, a single protein, N, can
perform multiple functions in the same cytoplasm. I uncovered the mechanism by which SARS-CoV-2 N
recognizes RNA to undergo condensation. Now, I am interested in understanding how the interaction of N with
viral RNA controls condensation and impacts important viral functions. RESEARCH: In my K99/R00 research;
(Aim 1) I will unbiasedly probe RNA-RNA interactions to understand how RNA is arranged and how
interactions are modulated by condensation for genome circularization. (Aim 2) I will develop BSL2 assays to
determine which viral processes require N condensation. (Aim 3) I will design synthetic RNA-binding proteins
and small molecules to disrupt N condensation. TRAINING: I will complete my training period in Dr. Amy
Gladfelter’s lab. During the training period, I will further develop my skills in RNA structure probing SHAPE
(RNA structure), RNP-map (RNA/protein crosslinking), and SHAPE-Jump (RNA-RNA crosslinking) in
collaboration with Weeks lab. I will learn how to design synthetic IDPs for the creation of bespoke RNA-binding
proteins (Chilkoti). I will screen an existing small molecule library to identify inhibitors of N condensation
(Fiorti). I will learn how to culture and genetically manipulate a BSL2 model betacoronavirus, MHV (Sheahan
and Cameron). ENVIRONMENT: Dr. Amy Gladfelter is one of the world’s leading experts on the role of RNA in
condensation. Additionally, UNC has some of the world’s leading experts in coronavirus biology including labs
with BSL3 facilities offering the possibility to test newly developed N condensation inhibitors with live virus. I
plan to further my growth through participation in RNA and condensation conferences, and attendance of
courses in virology. I also plan on using my K99/R00 to ease my transition into running my own lab as an
independent investigator. I am particularly interested in joining a university with strong biochemistry, molecular
biology, virology, and biomedical engineering departments who are looking for an RNA structure enthusiast to
join their ranks! IMPACT ON PUBLIC HEALTH: This work will advance our understanding of how
betacoronavirus N recognize viral RNA sequences and may one day lead to novel therapeutics.
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