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

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    Type
    Article
    Authors
    Li, Lixin
    Atef, Ahmed
    Piatek, Agnieszka Anna cc
    Ali, Zahir cc
    Piatek, Marek J. cc
    Aouida, Mustapha cc
    Sharakuu, Altanbadralt
    Mahjoub, Ali cc
    Wang, Guangchao
    Khan, Mohammad Suhail
    Fedoroff, Nina V. cc
    Zhu, Jian-Kang cc
    Mahfouz, Magdy M. cc
    KAUST Department
    Advanced Nanofabrication, Imaging and Characterization Core Lab
    Biological and Environmental Science and Engineering (BESE) Division
    Bioscience Program
    Center for Desert Agriculture
    Core Labs
    Imaging and Characterization Core Lab
    Laboratory for Genome Engineering
    Plant Science
    Plant Science Program
    Date
    2013-07
    Permanent link to this record
    http://hdl.handle.net/10754/562847
    
    Metadata
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    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. © 2013 The Author.
    Citation
    Li, L., Atef, A., Piatek, A., Ali, Z., Piatek, M., Aouida, M., … Mahfouz, M. M. (2013). Characterization and DNA-Binding Specificities of Ralstonia TAL-Like Effectors. Molecular Plant, 6(4), 1318–1330. doi:10.1093/mp/sst006
    Sponsors
    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.
    Publisher
    Elsevier BV
    Journal
    Molecular Plant
    DOI
    10.1093/mp/sst006
    PubMed ID
    23300258
    PubMed Central ID
    PMC3716395
    Additional Links
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3716395
    ae974a485f413a2113503eed53cd6c53
    10.1093/mp/sst006
    Scopus Count
    Collections
    Articles; Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; Imaging and Characterization Core Lab; Plant Science Program; Center for Desert Agriculture

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