Loss of ATM function enhances recombinant adeno-associated virus transduction and integration through pathways similar to UV irradiation.
Ataxia telangiectasia is caused by a genetic defect in the ATM gene that results in altered cellular sensitivity to DNA-damaging agents such as gamma-irradiation. ATM deficiency is associated with an increased incidence of neurological disorders, immune deficiency, and cancer. In this report we demonstrate that recombinant adeno-associated virus (rAAV) gene transfer in ATM-deficient fibroblasts is significantly enhanced over normal fibroblast cell lines. This enhancement of rAAV transduction in AT cells is correlated with an increased abundance of circular form rAAV genomes, as well as a higher number of integrated head-to-tail concatamer proviral genomes. Studies evaluating AAV trafficking using Cy3-labeled virus suggest that a nuclear mechanism is responsible for increased rAAV transduction in AT cells, because binding, endocytosis, and nuclear trafficking of virus are unaffected by the AT phenotype. Additionally, the profile of rAAV transduction after UV irradiation is significantly blunted in AT cells, suggesting that the level of DNA repair enzymes normally associated with UV augmentation of viral transduction may already be maximally elevated. These results further expand our understanding of genes involved in rAAV transduction.
Sanlioglu
S
S
Department of Anatomy, University of Iowa School of Medicine, Iowa City, Iowa, 52242, USA.
Benson
P
P
Engelhardt
J F
JF
D001260:Ataxia Telangiectasia / Q000235:genetics; D064007:Ataxia Telangiectasia Mutated Proteins; D015139:Blotting, Southern; D018797:Cell Cycle Proteins; D002460:Cell Line; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D004268:DNA-Binding Proteins; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology*; D005347:Fibroblasts / Q000528:radiation effects; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D025521:Tumor Suppressor Proteins; D014466:Ultraviolet Rays; D016662:Virus Integration
D018797:Cell Cycle Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D025521:Tumor Suppressor Proteins; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases