Host Cell DNA Repair Pathways in Adeno-Associated Viral Genome Processing
| Autor: | Vivian W. Choi, Douglas M. McCarty, R. Jude Samulski |
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| Rok vydání: | 2006 |
| Předmět: |
Male
DNA Repair DNA repair RecQ helicase Genetic Vectors Immunology Mice Transgenic Genome Viral Mice SCID Biology Microbiology Genome Cell Line law.invention Mice chemistry.chemical_compound Plasmid DNA Maintenance Viral life cycle law Virology Animals Humans DNA Breaks Double-Stranded DNA Primers Genetics Base Sequence Dependovirus Virus-Cell Interactions Cell biology Mice Inbred C57BL chemistry Insect Science DNA Viral Recombinant DNA Female DNA Plasmids |
| Zdroj: | Journal of Virology. 80:10346-10356 |
| ISSN: | 1098-5514 0022-538X |
| Popis: | Recentstudies have shown that wild-type and recombinant adeno-associated virus (AAV and rAAV) genomes persist in human tissue predominantly as double-stranded (ds) circular episomes derived from input linear single-stranded virion DNA. Using self-complementary recombinant AAV (scAAV) vectors, we generated intermediates that directly transition to ds circular episomes. The scAAV genome ends are palindromic hairpin-structured terminal repeats, resembling a double-stranded break repair intermediate. Utilizing this substrate, we found cellular DNA recombination and repair factors to be essential for generating circular episomal products. To identify the specific cellular proteins involved, the scAAV circularization-dependent vector was used as a reporter in 19 mammalian DNA repair-deficient cell lines. The results show that RecQ helicase family members (BLM and WRN), Mre11 and NBS1 of the Mre11-Rad50-Nbs1 (MRN) complex, and ATM are required for efficient scAAV genome circularization. We further demonstrated that the scAAV genome requires ATM and DNA-PK CS , but not NBS1, to efficiently convert to a circular form in nondividing cells in vivo using transgenic mice. These studies identify specific pathways involved for further elucidating viral and cellular mechanisms of DNA maintenance important to the viral life cycle and vector utilizations. |
| Databáze: | OpenAIRE |
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