Inhibition of Binding of Tomato Yellow Leaf Curl Virus Rep to its Replication Origin by Artificial Zinc-Finger Protein
- 11 May 2012
- journal article
- research article
- Published by Springer Science and Business Media LLC in Molecular Biotechnology
- Vol. 54 (2), 198-203
- https://doi.org/10.1007/s12033-012-9552-5
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.Keywords
This publication has 14 references indexed in Scilit:
- Molecular Biodiversity, Taxonomy, and Nomenclature of Tomato Yellow Leaf Curl-like VirusesPublished by Springer Science and Business Media LLC ,2007
- Gene silencing of Tomato Yellow Leaf Curl VirusPublished by Springer Science and Business Media LLC ,2007
- Inhibition of Virus DNA Replication by Artificial Zinc Finger ProteinsJournal of Virology, 2005
- Rational Design of Artificial Zinc-Finger Proteins Using a Nondegenerate Recognition Code TableBiochemistry, 2002
- The Multifunctional Character of a Geminivirus Replication Protein Is Reflected by Its Complex Oligomerization PropertiesJournal of Biological Chemistry, 2000
- Geminiviruses: models for plant DNA replication, transcription, and cell cycle regulation.2000
- A 2.2 Å resolution crystal structure of a designed zinc finger protein bound to DNANature Structural & Molecular Biology, 1996
- Agroinoculation of Tomato Yellow Leaf Curl Virus (TYLCV) Overcomes the Virus Resistance of Wild Lycopersicon SpeciesPlant Breeding, 1994
- Geminivirus replication origins have a modular organization.Plant Cell, 1994
- A geminivirus replication protein is a sequence-specific DNA binding protein.Plant Cell, 1992