Manipulation de protéines à bromodomaine d'Arabidopsis par une acetyltransférase de la famille YopJ de chez Ralstonia solanacearum
Bibliographic record
Abstract
Epigenetic mechanisms contribute to the regulation of gene expression without changing its sequence by influencing the chromatin structure. Increasing evidence reveal that pathogens display virulence strategies that can interfere with host epigenetic mechanisms. This is particularly well described in animal pathogens but less in plant pathogens. Especially, very few evidence relate such virulence strategies used by plant pathogenic bacteria. Ralstonia solanacearum is the causal agent of the bacterial wilt disease, which can affect more than 250 plant species including major crops and model plants such as Arabidopsis thaliana. As a potent R. solanacearum virulence factor, PopP2 is an acetyltransferase from the YopJ family that dampens basal immune responses by targeting defensive WRKY transcription factors. In order to better understand the virulence activities of PopP2, we searched for PopP2-interacting proteins using a yeast two hybrid assay, and identified the GTE9 and GTE11 proteins from the GTE family (General Transcription factor, group E). GTE proteins possess a bromodomain, a specific protein module allowing interaction with acetylated lysine residues, notably on histones tails suggesting that they could be involved in epigenetic-related processes. GTE9 and GTE11 were previously shown to (i) co-localise and interact with PopP2 in the plant nucleus, and (ii) to be acetylated by PopP2. Also, GTE9 and GTE11 were shown to interact in planta with Histone H4 through their bromodomain, suggesting that they function as epigenetic readers whose manipulation by PopP2 would promote R. solanacearum virulence. In this context, the main objectives of my PhD were to better understand the the function of GTE9 and GTE11, by trying to determine how they can be manipulated by PopP2 and whether these proteins play a role in the plant response to R. solanacearum. Mass-spectrometry-based analysis enabled us to map the lysine residues modified by PopP2 in GTE9 and GTE11. Several of these residues are conserved between the two proteins and localised on either side of their bromodomain. By semi-quantitative FRET-FLIM assay performed in vivo, we demonstrated that GTE9 interaction with Histone H4 is altered by PopP2 acetyltransferase activity suggesting that PopP2 uses acetylation to dissociate GTE9 from chromatin. In addition to GTE9 and GTE11, PopP2 acetylates several other GTE members. Regarding the role of GTE9 and GTE11 in the plant response to R. solanacearum, GTE9 and GTE11 over-expressing lines displayed an enhanced disease response to R. solanacearum that depended on PopP2 enzymatic activity. Overall, these data indicate that GTE9 and GTE11 behave as epigenetic readers that are manipulated by a plant bacterial pathogen through their targeting by a YopJ family acetyltransferase. GTE proteins could represent key virulence targets for R. solanacearum since PopP1, an additional YopJ family acetyltransferase that belongs to its effector repertoire, also interacts with several of these proteins. How the targeting of GTE proteins is mechanistically impacting the overall course of R. solanacearum infection remains elusive. To answer this question, we undertook a ChIP-seq analysis aimed at identifying the chromatin regions targeted by GTE9 and GTE11 (approach in progress). In addition to this approach, we wanted to identify more globally the chromatin sites visited by PopP2 in Arabidopsis. For this, we have initiated a second ChIP-seq analysis using various molecular tools including tagged versions of PopP2 for in planta delivery through a bacterial type III secretion system. Overall, this PhD work allows to progress on the understanding of a virulence strategy used by a plant bacterial pathogen that consist in manipulating host epigenetic components to promote infection.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".