TY - JOUR
T1 - Characterization and DNA-binding specificities of Ralstonia TAL-like effectors
AU - Li, Lixin
AU - Atef, Ahmed
AU - Piatek, Agnieszka Anna
AU - Ali, Zahir
AU - Piatek, Marek J.
AU - Aouida, Mustapha
AU - Sharakuu, Altanbadralt
AU - Mahjoub, Ali
AU - Wang, Guangchao
AU - Khan, Mohammad Suhail
AU - Fedoroff, Nina V.
AU - Zhu, Jian-Kang
AU - Mahfouz, Magdy M.
N1 - 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.
PY - 2013/7
Y1 - 2013/7
N2 - 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.
AB - 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.
UR - http://hdl.handle.net/10754/562847
UR - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3716395
UR - http://www.scopus.com/inward/record.url?scp=84880287458&partnerID=8YFLogxK
U2 - 10.1093/mp/sst006
DO - 10.1093/mp/sst006
M3 - Article
C2 - 23300258
SN - 1674-2052
VL - 6
SP - 1318
EP - 1330
JO - Molecular Plant
JF - Molecular Plant
IS - 4
ER -