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Characterization and DNA-Binding Specificities of Ralstonia TAL-Like Effectors.

Bibliography:

Li, L., A. Atef, A. Piatek, Z. Ali, M. Piatek, M. Aouida​, A. Sharakuu, A. Mahjoub, G. Wang, S. Khan, N. V. Fedoroff, J. K. Zhu and Magdy M. Mahfouz (2013). "Characterization and DNA-Binding Specificities of Ralstonia TAL-Like Effectors." Molecular Plant. (4):1318-30.​​

Authors:

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

Keywords:

Ralstonia solanacearum, TAL effectors, TALE activators and repressors, TALE nucleases (TALENs), genome engineering, targeted genome modifications

Year:

2013

Abstract:

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.

ISSN:

1674-2052