Muramatsu S, Handa A, Kajigaya S, Brown K E
Hematology Branch, National Heart, Lung and Blood Institute, Bethesda, MD 20892-1652, USA.
J Gen Virol. 1998 Sep;79 ( Pt 9):2157-61. doi: 10.1099/0022-1317-79-9-2157.
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.
虽然腺相关病毒（AAV）复制需要Rep蛋白，但对于与Rep相互作用的细胞蛋白却知之甚少。我们在此证明，与Gal4激活域融合的转录阳性辅因子4（PC4，p15）与与Gal4 DNA结合域融合的AAV-2和AAV-3 Rep蛋白相互作用，导致酵母双杂交系统中的报告基因激活。除了其共激活功能外，PC4最近还被证明参与猿猴病毒40的复制。为了研究PC4-Rep蛋白相互作用的功能作用，将PC4表达质粒和AAV-3的感染性克隆共转染到293-31细胞中，随后用辅助腺病毒进行超感染。在转染PC4的细胞中拯救出的AAV-3基因组数量显著增加。我们的结果支持PC4可能参与AAV复制，并可用于高效生产用于基因治疗的AAV载体。
