Adeno-Associated Virus Rep Represses the Human Integration Site Promoter by Two Pathways That Are Similar to Those Required for the Regulation of the Viral p5 Promoter
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Cell lines and viruses.
Infections.
Reporter constructs.
Rep-expressing constructs.
Transient transfections.
Plasmid uptake determination.
Real-time qRT-PCR.
Northern blot analysis.
Western blot analysis.
Immunofluorescence.
Fluorescence-activated cell sorting.
RESULTS
Under permissive conditions, AAV2 infection leads to downregulation of PPP1R12C expression.
Both large and small Rep proteins repress PPP1R12C promoter activity.
The NTP-binding motif is required for Rep52- and Rep40-mediated repression of the p5 and PPP1R12C promoters.
The RBS and the NTP-binding motif are both required for Rep78/Rep68-mediated repression of the p5 and PPP1R12C promoters.
Rep-mediated repression of the antisense promoter requires only a functional NTP-binding motif.
DISCUSSION
ACKNOWLEDGMENTS
REFERENCES
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