Characterization and DNA-binding specificities of Ralstonia TAL-like effectors
Piatek, Agnieszka Anna
Piatek, Marek J.
Khan, Mohammad Suhail
Fedoroff, Nina V.
Mahfouz, Magdy M.
KAUST DepartmentAdvanced Nanofabrication, Imaging and Characterization Core Lab
Biological and Environmental Sciences and Engineering (BESE) Division
Desert Agriculture Initiative
Imaging and Characterization Core Lab
Laboratory for Genome Engineering
Plant Science Program
Permanent link to this recordhttp://hdl.handle.net/10754/562847
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AbstractTranscription 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.
SponsorsThis 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.
PubMed Central IDPMC3716395
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