Inhibition of DNA replication of human papillomavirus by artificial zinc finger proteins.
Recently, we demonstrated that plant DNA virus replication was inhibited in planta by using an artificial zinc finger protein (AZP) and created AZP-based transgenic plants resistant to DNA virus infection. Here we apply the AZP technology to the inhibition of replication of a mammalian DNA virus, human papillomavirus type 18 (HPV-18). Two AZPs, designated AZP(HPV)-1 and AZP(HPV)-2, were designed by using our nondegenerate recognition code table and were constructed to block binding of the HPV-18 E2 replication protein to the replication origin. Both of the newly designed AZPs had much higher affinities towards the replication origin than did the E2 protein, and they efficiently blocked E2 binding in vitro. In transient replication assays, both AZPs inhibited viral DNA replication, especially AZP(HPV)-2, which reduced the replication level to approximately 10%. We also demonstrated in transient replication assays, using plasmids with mutant replication origins, that AZP(HPV)-2 could precisely recognize the replication origin in mammalian cells. Thus, it was demonstrated that the AZP technology could be applied not only to plant DNA viruses but also to mammalian DNA viruses.
Mino
Takashi
T
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-Ku, Kyoto 615-8510, Japan.
Hatono
Takeaki
T
Matsumoto
Naoki
N
Mori
Tomoaki
T
Mineta
Yusuke
Y
Aoyama
Yasuhiro
Y
Sera
Takashi
T
D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis*; D016679:Genome, Viral; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
D004279:DNA, Viral