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

Handle URI:
http://hdl.handle.net/10754/562847
Title:
Characterization and DNA-binding specificities of Ralstonia TAL-like effectors
Authors:
Li, Lixin; Atef, Ahmed; Piatek, Agnieszka Anna ( 0000-0001-9587-6360 ) ; Ali, Zahir ( 0000-0002-7814-8908 ) ; Piatek, Marek J.; Aouida, Mustapha ( 0000-0001-7945-5320 ) ; Sharakuu, Altanbadralt; Mahjoub, Ali ( 0000-0002-9510-3218 ) ; Wang, Guangchao; Khan, Mohammad Suhail; Fedoroff, Nina V.; Zhu, Jiankang; Mahfouz, Magdy M. ( 0000-0002-0616-6365 )
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.
KAUST Department:
Center for Desert Agriculture; Advanced Nanofabrication, Imaging and Characterization Core Lab; Bioscience Program; Core Labs; Biological and Environmental Sciences and Engineering (BESE) Division
Publisher:
Elsevier BV
Journal:
Molecular Plant
Issue Date:
Jul-2013
DOI:
10.1093/mp/sst006
PubMed ID:
23300258
PubMed Central ID:
PMC3716395
Type:
Article
ISSN:
16742052
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.
Additional Links:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3716395
Appears in Collections:
Articles; Bioscience Program; Advanced Nanofabrication, Imaging and Characterization Core Lab; Center for Desert Agriculture; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorLi, Lixinen
dc.contributor.authorAtef, Ahmeden
dc.contributor.authorPiatek, Agnieszka Annaen
dc.contributor.authorAli, Zahiren
dc.contributor.authorPiatek, Marek J.en
dc.contributor.authorAouida, Mustaphaen
dc.contributor.authorSharakuu, Altanbadralten
dc.contributor.authorMahjoub, Alien
dc.contributor.authorWang, Guangchaoen
dc.contributor.authorKhan, Mohammad Suhailen
dc.contributor.authorFedoroff, Nina V.en
dc.contributor.authorZhu, Jiankangen
dc.contributor.authorMahfouz, Magdy M.en
dc.date.accessioned2015-08-03T11:12:33Zen
dc.date.available2015-08-03T11:12:33Zen
dc.date.issued2013-07en
dc.identifier.issn16742052en
dc.identifier.pmid23300258en
dc.identifier.doi10.1093/mp/sst006en
dc.identifier.urihttp://hdl.handle.net/10754/562847en
dc.description.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.en
dc.description.sponsorshipThis 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.en
dc.publisherElsevier BVen
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3716395en
dc.subjectgenome engineeringen
dc.subjectRalstonia solanacearumen
dc.subjectTAL effectorsen
dc.subjectTALE activators and repressorsen
dc.subjectTALE nucleases (TALENs)en
dc.subjecttargeted genome modificationsen
dc.titleCharacterization and DNA-binding specificities of Ralstonia TAL-like effectorsen
dc.typeArticleen
dc.contributor.departmentCenter for Desert Agricultureen
dc.contributor.departmentAdvanced Nanofabrication, Imaging and Characterization Core Laben
dc.contributor.departmentBioscience Programen
dc.contributor.departmentCore Labsen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalMolecular Planten
dc.identifier.pmcidPMC3716395en
dc.contributor.institutionDepartment of Biological Sciences, Faculty of Sciences, King Abdulaziz University, PO Box 80203, Jeddah 21589, Saudi Arabiaen
dc.contributor.institutionDepartment of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, United Statesen
kaust.authorLi, Lixinen
kaust.authorAouida, Mustaphaen
kaust.authorMahjoub, Alien
kaust.authorWang, Guangchaoen
kaust.authorFedoroff, Nina V.en
kaust.authorMahfouz, Magdy M.en
kaust.authorPiatek, Agnieszka Annaen
kaust.authorPiatek, Marek J.en
kaust.authorSharakuu, Altanbadralten
kaust.authorKhan, Mohammad Suhailen

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