Biology subjects

Erkes, A.

Publications and source records attributed to Erkes, A..

4 recordsLinked to original sources

bioRxiv2024

Time-resolved RNA-seq data reveal dynamic expressional behaviour of TALE target genes

Xanthomonas oryzae bacteria infect rice (Oryza sativa) plants causing substantial harvest loss. During the infection, the bacteria translocate a collection of effector proteins into the host plant cells. This includes transcription activator-like effectors (TALEs) that act as transcriptional activators of plant genes. To understand the dynamics of TALE action during the infection, we collected RNA-seq data at different time points after the infection of rice plants with two representative pathogens of the two rice-pathogenic Xanthomonas oryzae ssp., namely Xan-thomonas oryzae pv. oryzae PXO83 and Xanthomonas oryzae pv. oryzicola BAI35. We observed that the induction of direct TALE target transcripts already starts 24h after the infection and is widely established after 36h, while 72h after the infection, secondary targets and downstream effects start to dominate the set of differentially expressed transcripts relative to control. Based on computational predictions of TALE targets combined with expression data, we established criteria that may help to identify direct TALE targets, which are related to expression dynamics but also shifted transcription start sites, and compiled a list of high-confidence TALE targets. Using genome-wide target predictions, we further discovered several non-coding and anti-sense transcripts that are likely induced by TALEs. Finally, we investigated different strategies to link putative secondary targets to transcription factors that are induced by TALEs during the infection.

plant biology↗

bioRxiv2022

Assembling highly repetitive Xanthomonas TALomes using Oxford Nanopore sequencing

Most plant-pathogenic Xanthomonas bacteria harbor transcription activator-like effector (TALE) genes, which function as transcriptional activators of host plant genes and support infection. The entire repertoire of up to 29 TALE genes of a Xanthomonas strain is also referred to as TALome. The DNA-binding domain of TALEs is comprised of highly conserved repeats and TALE genes often occur in gene clusters, which precludes the assembly of TALE-carrying Xanthomonas genomes based on standard sequencing approaches. Here, we report the successful assembly of the 5 Mbp genomes of five Xanthomonas strains from Oxford Nanopore Technologies (ONT) sequencing data. For one of these strains, Xanthomonas oryzae pv. oryzae (Xoo) PXO35, we illustrate why Illumina short reads and longer PacBio reads are insufficient to fully resolve the genome. While ONT reads are perfectly suited to yield highly contiguous genomes, they suffer from a specific error profile within homopolymers. To still yield complete and correct TALomes from ONT assemblies, we present a computational correction pipeline specifically tailored to TALE genes, which yields at least comparable accuracy as Illumina-based polishing. We further systematically assess the ONT-based pipeline for its multiplexing capacity and find that, combined with computational correction, the complete TALome of Xoo PXO35 could have been reconstructed from less than 20,000 ONT reads. Our results indicate that multiplexed ONT sequencing combined with a computational correction of TALE genes constitutes a highly capable tool for characterizing the TALomes of huge collections of Xanthomonas strains in the future.

bioinformatics↗

bioRxiv2021

Epigenetic features improve TALE target prediction

The yield of many crop plants can be substantially reduced by plant-pathogenic Xanthomonas bacteria. The infection strategy of many Xanthomonas strains is based on transcription activator-like effectors (TALEs), which are secreted into the host cells and act as transcriptional activators of plant genes that are beneficial for the bacteria. The modular DNA binding domain of TALEs contains tandem repeats, each comprising two hyper-variable amino acids. These repeat-variable diresidues (RVDs) bind to a continuous DNA stretch (a target box) and determine the specificity of a TALE. All available tools for the prediction of TALE targets within the host plant suffer from many false positives. In this paper we propose a strategy to improve prediction accuracy by considering the epigenetic state of the host plant genome in the region of the target box. To this end, we extend our previously published tool PrediTALE by two epigenetic features: (i) We allow for filtering target boxes according to chromatin accessibility and (ii) we allow for considering the methylation state of cytosines within the target box during prediction, since DNA methylation may affect the binding specificity of RVDs. Here, we determine the epigenetic features from publicly available DNase-seq, ATAC-seq, and WGBS-seq data in rice. We benchmark the utility of both epigenetic features separately and in combination, deriving ground-truth from RNA-seq infections studies in rice. We find an improvement for each individual epigenetic feature, but especially the combination of both. Having established an advantage in TALE target predicting considering epigenetic features, we use these data for promoterome and genome-wide scans by our new tool EpiTALE, leading to several novel putative virulence targets. Our results suggest that it would be worthwhile to collect condition-specific chromatin accessibility data and methylation information when studying putative virulence targets of Xan-thomonas TALEs.

bioinformatics↗

bioRxiv2019

PrediTALE: A novel model learned from quantitative data allows for new perspectives on TALE targeting

Plant-pathogenic Xanthomonas bacteria secret transcription activator-like effectors (TALEs) into host cells, where they act as transcriptional activators on plant target genes to support bacterial virulence. TALEs have a unique modular DNA-binding domain composed of tandem repeats. Two amino acids within each tandem repeat, termed repeat-variable diresidues, bind to contiguous nucleotides on the DNA sequence and determine target specificity.\n\nIn this paper, we propose a novel approach for TALE target prediction to identify potential virulence targets. Our approach accounts for recent findings concerning TALE targeting, including frame-shift binding by repeats of aberrant lengths, and the flexible strand orientation of target boxes relative to the transcription start of the downstream target gene. The computational model can account for dependencies between adjacent RVD positions. Model parameters are learned from the wealth of quantitative data that have been generated over the last years.\n\nWe benchmark the novel approach, termed PrediTALE, using RNA-seq data after Xanthomonas infection in rice, and find an overall improvement of prediction performance compared with previous approaches. Using PrediTALE, we are able to predict several novel putative virulence targets. However, we also observe that no target genes are predicted by any prediction tool for several TALEs, which we term orphan TALEs for this reason. We postulate that one explanation for orphan TALEs are incomplete gene annotations and, hence, propose to replace promoterome-wide by genome-wide scans for target boxes. We demonstrate that known targets from promoterome-wide scans may be recovered by genome-wide scans, whereas the latter, combined with RNA-seq data, are able to detect putative targets independent of existing gene annotations.\n\nAuthor summaryDiseases caused by plant-pathogenic Xanthomonas bacteria are a serious threat for many important crop plants including rice. Efficiently protecting plants from these pathogens requires a deeper understanding of infection strategies. For many Xanthomonas strains, such infection strategies depend on a special class of effector proteins, termed transcription activator-like effectors (TALEs). TALEs may specifically activate genes of the host plant and, by this means, re-program the plant cell for the benefit of the pathogen. Target sequences and, consequently, target genes of a specific TALE may be predicted computationally from its amino acids. Here, we propose a novel approach for TALE target prediction that makes use of several insights into TALE biology but also of broad experimental data gained over the last years. We demonstrate that this approach yields a higher prediction accuracy than previous approaches. We further postulate that a strategy change from a restricted search only considering promoters of annotated genes to a broad genome-wide search is feasible and yields novel targets including previously neglected protein-coding genes but also non-coding RNAs of possibly regulatory function.

bioinformatics↗