Belmont University Research Symposium (BURS)

Publication Date

Spring 4-19-2023

College

Sciences and Mathematics, College of

Department

Biology, Department of

BURS Faculty Advisor

Rebecca Adams

Presentation Type

Poster Presentation

Abstract

Project Title: Photoactivatable Probes Reveal Structural Information Between InfluenzaA NP1 and Host mRNA Export Factors.

The Influenza virus is known to cause a respiratory illness and seasonal flu epidemics. One of the influenza genes encodes a viral nucleoprotein (NP) that binds to the viral RNA genome and is required for viral replication and transcription. The viral NP protein hijacks the host mRNA export factor UAP56 in order to properly package and export the viral RNA. However, the exact mechanism of the interaction between NP and UAP56 is still unclear. This study aims explore the UAP56 residues necessary for interaction with NP by utilizing the photo crosslinking approach. To do this, a photoreactive, unnatural amino acid, p-benzoyl-l-phenylalanine (pBpa), which crosslinks to any adjacent CH group when exposed to UV, was integrated into the newly synthesized protein by using Amber codon suppression. Through these methods, crosslinking between NP and UAP56 was observed only at specific residues. In the future, we anticipate that mass spectrometry of the crosslinking products to reveal mechanistic information between this interaction. Additionally, we plan to use negative stain electron microscopy (EM) and cryo-EM to obtain structural information of this interaction.

Key words: viral genome, mRNA export, photo crosslinking, protein residues.

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