RESUMO
The pathogenicity of many bacteria depends on the injection of effector proteins via type III secretion into eukaryotic cells in order to manipulate cellular processes. TAL (transcription activator-like) effectors from plant pathogenic Xanthomonas are important virulence factors that act as transcriptional activators in the plant cell nucleus, where they directly bind to DNA via a central domain of tandem repeats. Here, we show how target DNA specificity of TAL effectors is encoded. Two hypervariable amino acid residues in each repeat recognize one base pair in the target DNA. Recognition sequences of TAL effectors were predicted and experimentally confirmed. The modular protein architecture enabled the construction of artificial effectors with new specificities. Our study describes the functionality of a distinct type of DNA binding domain and allows the design of DNA binding domains for biotechnology.
Assuntos
Motivos de Aminoácidos , DNA de Plantas/química , DNA de Plantas/metabolismo , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Ativação Transcricional , Xanthomonas/metabolismo , Sequência de Aminoácidos , Arabidopsis/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Pareamento de Bases , Sequência de Bases , Biotecnologia , Capsicum/genética , Genes de Plantas , Modelos Biológicos , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Ligação Proteica , Sequências Repetitivas de Aminoácidos , Nicotiana/genética , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Xanthomonas/patogenicidadeRESUMO
SUMMARY Jasmonic acid and related oxylipin compounds are plant signalling molecules that are involved in the response to pathogens, insects, wounding and ozone. To explore further the role of jasmonates in stress signal transduction, the response of two jasmonate-signalling mutants, jin1 and jin4, to pathogens and ozone was analysed in this study. Upon treatment with the biotrophic bacterial pathogen Pseudomonas syringae, endogenous jasmonate levels increased in jin1 and jin4 similar to wild-type, demonstrating that these mutants are not defective in jasmonate biosynthesis. Jin1 but not jin4 is more resistant to P. syringae and this higher resistance is accompanied by higher levels of salicylic acid. Jin1 is also more resistant to the necrotrophic fungal pathogen Botrytis cinerea and shows wild-type sensitivity to ozone whereas jin4 is more susceptible to B. cinerea and ozone. These results indicate that the mutations in jin1 and jin4 affect different branches of the jasmonate signalling pathway. Additionally, in this combination of phenotypes, jin1 is unique among all other jasmonate-related mutants described thus far. These data also provide support for a crosstalk between the jasmonate and salicylate pathways.