title: Inhibition of binding of tomato yellow leaf curl virus rep to its replication origin by artificial zinc-finger protein.
abstract: Previously we demonstrated that inhibition of replication-associated protein (Rep) binding to its replication origin by artificial zinc-finger proteins (AZPs) is a powerful method to prevent plant virus infection in vivo. In the present study, we applied the AZP technology to Tomato yellow leaf curl virus (TYLCV), which is a limiting factor in tomato cultivation worldwide. First, we determined 5'-ATCGGTGT ATCGGTGT-3' in the 195-bp intergenic region of the TYLCV-Israel strain, a strain reported first among TYLCV strains, as the Rep-binding site by gel shift assays. We then constructed a 6-finger AZP that bound to a 19-bp DNA including the Rep-binding site. We demonstrated that the binding affinity of the AZP was >1,000-fold greater than that of Rep and that the AZP inhibited Rep binding completely in vitro. Because the binding capability of the AZP was same as that of the AZP previously designed for geminivirus-resistant Arabidopsis thaliana, we predict that the present AZP will prevent TYLCV infection in vivo.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Mori
authors.forename: Kosuke
authors.initials: K
authors.lastname: Takenaka
authors.forename: Fumiya
authors.initials: F
authors.lastname: Domoto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Aoyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Sera
mesh_terms: D053839:Begomovirus / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D018551:Solanum lycopersicum / Q000821:virology*; D011485:Protein Binding / Q000235:genetics*; D018741:Replication Origin / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins