Transcription-positive cofactor 4 enhances rescue of adeno-associated virus genome from an infectious clone.
While Rep proteins are required for adeno-associated virus (AAV) replication, little is known about cellular proteins that interact with Rep. We demonstrate here that transcription-positive cofactor 4 (PC4, p15) fused to Gal4-activating domain interacted with both AAV-2 and AAV-3 Rep proteins fused to Gal4 DNA-binding domain, leading to reporter activation in the yeast two-hybrid system. In addition to its coactivating function, PC4 recently has been shown to be involved in replication of simian virus 40. To study a functional role for the PC4-Rep protein interaction, 293-31 cells were cotransfected with a PC4 expression plasmid and an infectious clone of AAV-3, followed by super-infection with helper adenovirus. A significantly increased number of AAV-3 genomes were rescued in PC4 transfected cells. Our results support a possible involvement of PC4 in AAV replication and may be used in efficient production of AAV vectors for gene therapy.
Muramatsu
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S
Hematology Branch, National Heart, Lung and Blood Institute, Bethesda, MD 20892-1652, USA. muramats@nih.gov
Handa
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Kajigaya
S
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Brown
K E
KE
D000256:Adenoviridae / Q000502:physiology; D002460:Cell Line; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D016679:Genome, Viral; D006378:Helper Viruses / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C490043:IFRD2 protein, human; D017874:Immediate-Early Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C045264:replication initiator protein; C068734:rep proteins, Adeno-associated virus 2; D004265:DNA Helicases