Characterization and DNA-binding specificities of Ralstonia TAL-like effectors

Lixin Li, Ahmed Atef, Agnieszka Anna Piatek, Zahir Ali, Marek J. Piatek, Mustapha Aouida, Altanbadralt Sharakuu, Ali Mahjoub, Guangchao Wang, Mohammad Suhail Khan, Nina V. Fedoroff, Jian-Kang Zhu, Magdy M. Mahfouz

Research output: Contribution to journalArticlepeer-review

49 Scopus citations


Transcription activator-like effectors (TALEs) from Xanthomonas sp. have been used as customizable DNA-binding modules for genome-engineering applications. Ralstonia solanacearum TALE-like proteins (RTLs) exhibit similar structural features to TALEs, including a central DNA-binding domain composed of 35 amino acid-long repeats. Here, we characterize the RTLs and show that they localize in the plant cell nucleus, mediate DNA binding, and might function as transcriptional activators. RTLs have a unique DNA-binding architecture and are enriched in repeat variable di-residues (RVDs), which determine repeat DNA-binding specificities. We determined the DNA-binding specificities for the RVD sequences ND, HN, NP, and NT. The RVD ND mediates highly specific interactions with C nucleotide, HN interacts specifically with A and G nucleotides, and NP binds to C, A, and G nucleotides. Moreover, we developed a highly efficient repeat assembly approach for engineering RTL effectors. Taken together, our data demonstrate that RTLs are unique DNA-targeting modules that are excellent alternatives to be tailored to bind to user-selected DNA sequences for targeted genomic and epigenomic modifications. These findings will facilitate research concerning RTL molecular biology and RTL roles in the pathogenicity of Ralstonia spp. © 2013 The Author.
Original languageEnglish (US)
Pages (from-to)1318-1330
Number of pages13
JournalMolecular Plant
Issue number4
StatePublished - Jul 2013

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: This research is funded from the Center for Desert Agriculture baseline funding. The work in J.-K.Z.'s lab is supported by the National Institute of Health grant R01GM070795.

ASJC Scopus subject areas

  • Plant Science
  • Molecular Biology


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