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Targeting the PDZ-ligand domain of avian influenza A viruses for novel therapeutics
Institution: Baylor College of Medicine, Houston, TX
Principal Investigator: Andrew Rice, PhD
Co-Investigators: Shinji Makino, PhD - UTMB, Galveston, TX
Expected Product:Novel therapeutics for avian influenza H5N1 viruses.
Description: Highly virulent H5N1 strains of influenza A virus are currently circulating in a number of avian species in Asia, Europe, and Africa. A devastating pandemic is possible if these strains acquire the ability for efficient human-to-human spread. A recent study found that the viral NS1A protein is associated with the virulence of H5N1 viruses. It was also found that avian influenza NS1A proteins, as well as NS1A proteins from recent human H5N1 infections, contain a domain at their carboxyl termini termed the PDZ-ligand domain, and this domain associates in vitro with cellular PDZ proteins. PDZ proteins are a large class of proteins that are typically involved in cell-cell contract. It is therefore possible, if not likely, that the association of these NS1A proteins with key cellular PDZ proteins is an aspect of virulence in vivo. To develop an assay for therapeutics that target this viral function, two specific aims are proposed: (1) To develop a high-throughput screen for chemical inhibitors of the interaction between the NS1A PDZ-ligand domain and its cellular PDZ protein targets. This screen will be based upon a recently described method termed Reverse MAPPIT (mammalian protein-protein interaction trap). Once established, this system can be readily adapted for screens of inhibitors for other pathogens of relevance to the WCRE mission. (2) To identify cellular PDZ proteins that associate in vivo with the PDZ-ligand domain of the avian influenza virus NS1A protein. The identification of these cellular proteins will be accomplished by co-immunoprecipitation with the avian NS1 protein using a tandem-affinity purification method. A number of recent publications have reported success in screens
for small molecules that perturb protein-protein interactions.
These publications have therefore established the feasibility of
drugs that function by interfering with protein-protein interactions.
Completion of the work proposed here will provide an assay for
small molecules that block the interaction of the NS1A PDZ-ligand
domain with its targets, thus laying the foundation for development
of novel therapeutics for H5N1 infections.
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