Mutation in the platelet-derived growth factor receptor alpha inhibits adeno-associated virus type 5 transduction

Due to its non-pathogenic lifecycle, little is known about the cellular determinants of infection by adeno-associated virus (AAV). To identify these critical cellular factors, we took advantage of the gene transfer abilities of AAV in combination with a forward genetic selection to identify proteins critical for transduction by this virus. AAV serotype 5 (AAV5) vectors encoding the furin gene were used to transduce furin-deficient cells followed by selection with furin-dependent toxins. A population of cells specifically resistant to AAV5 transduction was identified and sequence analysis suggested all had a single amino acid mutation in the leader sequence of the platelet-derived growth factor receptor alpha (PDGFRα) gene. Characterization of this mutation suggested it inhibited PDGFRα trafficking resulting in limited expression on the plasma membrane. Mutagenesis and transfection experiments confirmed the effect of this mutation on PDGFRα trafficking, and the AAV5 resistant phenotype could be rescued by transfection with wild type PDGFRα.

► Forward genetic selection is a powerful tool for understanding complex host: virus interactions.
► A point mutation in the leader sequence of platelet-derived growth factor receptor alpha (PDGFRα) was found that blocks adeno-associated virus type 5 (AAV5) transduction.
► This finding suggests that PDGFRα is a critical protein in the transduction of AAV5.
► Virus entry is a rate-limiting step in the lifecycle of AAV5.