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Repeat 1 of TAL effectors affects target specificity for the base at position zero
AvrBs3, the founding member of the Xanthomonas transcription-activator-like effectors (TALEs), is translocated into the plant cell where it localizes to the nucleus and acts as transcription factor. The DNA-binding domain of AvrBs3 consists of 17.5 nearly-identical 34 amino acid-repeats. Each repeat...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4066769/ https://www.ncbi.nlm.nih.gov/pubmed/24792160 http://dx.doi.org/10.1093/nar/gku341 |
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author | Schreiber, Tom Bonas, Ulla |
author_facet | Schreiber, Tom Bonas, Ulla |
author_sort | Schreiber, Tom |
collection | PubMed |
description | AvrBs3, the founding member of the Xanthomonas transcription-activator-like effectors (TALEs), is translocated into the plant cell where it localizes to the nucleus and acts as transcription factor. The DNA-binding domain of AvrBs3 consists of 17.5 nearly-identical 34 amino acid-repeats. Each repeat specifies binding to one base in the target DNA via amino acid residues 12 and 13 termed repeat variable diresidue (RVD). Natural target sequences of TALEs are generally preceded by a thymine (T(0)), which is coordinated by a tryptophan residue (W232) in a degenerated repeat upstream of the canonical repeats. To investigate the necessity of T(0) and the conserved tryptophan for AvrBs3-mediated gene activation we tested TALE mutant derivatives on target sequences preceded by all possible four bases. In addition, we performed domain swaps with TalC from a rice pathogenic Xanthomonas because TalC lacks the tryptophan residue, and the TalC target sequence is preceded by cytosine. We show that T(0) works best and that T(0) specificity depends on the repeat number and overall RVD-composition. T(0) and W232 appear to be particularly important if the RVD of the first repeat is HD (‘rep1 effect’). Our findings provide novel insights into the mechanism of T(0) recognition by TALE proteins and are important for TALE-based biotechnological applications. |
format | Online Article Text |
id | pubmed-4066769 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-40667692014-06-24 Repeat 1 of TAL effectors affects target specificity for the base at position zero Schreiber, Tom Bonas, Ulla Nucleic Acids Res Molecular Biology AvrBs3, the founding member of the Xanthomonas transcription-activator-like effectors (TALEs), is translocated into the plant cell where it localizes to the nucleus and acts as transcription factor. The DNA-binding domain of AvrBs3 consists of 17.5 nearly-identical 34 amino acid-repeats. Each repeat specifies binding to one base in the target DNA via amino acid residues 12 and 13 termed repeat variable diresidue (RVD). Natural target sequences of TALEs are generally preceded by a thymine (T(0)), which is coordinated by a tryptophan residue (W232) in a degenerated repeat upstream of the canonical repeats. To investigate the necessity of T(0) and the conserved tryptophan for AvrBs3-mediated gene activation we tested TALE mutant derivatives on target sequences preceded by all possible four bases. In addition, we performed domain swaps with TalC from a rice pathogenic Xanthomonas because TalC lacks the tryptophan residue, and the TalC target sequence is preceded by cytosine. We show that T(0) works best and that T(0) specificity depends on the repeat number and overall RVD-composition. T(0) and W232 appear to be particularly important if the RVD of the first repeat is HD (‘rep1 effect’). Our findings provide novel insights into the mechanism of T(0) recognition by TALE proteins and are important for TALE-based biotechnological applications. Oxford University Press 2014-07-01 2014-05-03 /pmc/articles/PMC4066769/ /pubmed/24792160 http://dx.doi.org/10.1093/nar/gku341 Text en © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Molecular Biology Schreiber, Tom Bonas, Ulla Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title | Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title_full | Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title_fullStr | Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title_full_unstemmed | Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title_short | Repeat 1 of TAL effectors affects target specificity for the base at position zero |
title_sort | repeat 1 of tal effectors affects target specificity for the base at position zero |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4066769/ https://www.ncbi.nlm.nih.gov/pubmed/24792160 http://dx.doi.org/10.1093/nar/gku341 |
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