Inhibition of binding of tomato yellow leaf curl virus rep to its replication origin by artificial zinc-finger protein

Tomoaki Mori, Kosuke Takenaka, Fumiya Domoto, Yasuhiro Aoyama, Takashi Sera

Research output: Contribution to journalArticle

19 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)198-203
Number of pages6
JournalMolecular Biotechnology
Volume54
Issue number2
DOIs
Publication statusPublished - Jun 2013

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Keywords

  • Artificial zinc-finger protein
  • Common region
  • Geminivirus
  • Replication-associated protein
  • Tomato yellow leaf curl virus
  • Virus replication

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Molecular Biology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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