Biology subjects

Heintz, C.

Publications and source records attributed to Heintz, C..

8 recordsLinked to original sources

bioRxiv2025

Welcome pathogens: transient heat dampens the responses to acibenzolar-S-methyl beyond defenses in apple plants

Climate change affects plant-pathogen interactions, with disease outcome varying depending on pathosystem and environmental scenario. In Arabidopsis, a thermo-sensitive module of salicylic acid (SA) signaling makes immunity vulnerable to heat. The potent resistance inducer acibenzolar-S-methyl (ASM), an SA analogue that up-regulates transcription of defense genes, could restore plant protection under heat but not core SA signaling. Here, we investigated how high temperature rewires the ASM-induced responses of the apple immune system. We treated apple plants with ASM under contrasting heatwave scenarios and subsequently exposed them to Erwinia amylovora (the fire blight bacterium) or Venturia inaequalis (the apple scab fungus) while monitoring gene expression. While pre-exposing apple plants to high temperature did not change their susceptibility to pathogens, it drove a loss of ASM-induced protection. Transcriptomic analysis revealed broad dampening of ASM-regulation upon high temperature, for a wide range of biological processes beyond defense. We uncovered thermo-sensitive "resistance" and "susceptibility" marker genes with ASM-responsiveness being critically vulnerable to heat. We concluded that exposure to heatwave prevents ASM from fully mounting its protective responses in apple, not only lowering defenses but also offering more favorable hosting conditions. Our work highlights plant immunity as the joint outcome of resistant and susceptible responses. Summary StatementWe found that heatwaves "disarm" apples ability to mount an effective inducible-immunity response against two major diseases, fire blight and apple scab. Heatwaves not only prevent full expression of plant defenses, but also favor a physiological status that is beneficial to the pathogens.

plant biology↗

bioRxiv2024

Parallel in-depth analysis of repeat expansions: an updated Clin-CATS workflow for nanopore R10 flow cells

Hereditary ataxias, caused by expansions of short tandem repeats, are difficult to diagnose using traditional PCR and Southern blot methods, which struggle to detect complex repeat expansions and cannot assess repeat interruptions or methylation. We present an updated Clinical Nanopore Cas9-Targeted Sequencing (Clin-CATS) workflow for analyzing repeat expansions, now compatible with the Oxford Nanopore Technologies R10 flow cell. The workflow incorporates the ONT wf-human-variation Epi2Me workflow, including the Straglr tool to analyze basecalled reads, ensuring compatibility with past, current, and future sequencing chemistries. It expands the number of genes analyzed from 10 to 27 within the ataxia panel and introduces new gene panels for myopathy, neurodegeneration, and ALS/motor neuron disease. Validated with Coriell reference and clinical samples, this method improves the analysis of pathogenic repeat expansions, providing deeper insights into repeat structures while addressing the limitations of traditional approaches.

genetics↗

bioRxiv2024

Dual NBT Approach: Gene Editing and Intragenesis to promote Fire Blight Resistance in Apple using MdAGG defense lectins.

Fire blight, caused by the bacterium Erwinia amylovora, represents a significant threat to apple (Malus domestica) production. Currently, only a limited number of genes effectively involved in resistance to E. amylovora have been identified. Seeking new resistance candidates, we focused on a multigene family encoding amaranthin-like lectins, which are highly upregulated following chemical elicitation by acibenzolar-S-methyl (ASM). These lectins are believed to contribute to downstream defense by promoting bacterial aggregation, which led to their designation as M. domestica agglutinins (MdAGG). When loss-of-function editions were introduced into MdAGG genes, the plants ability to mount a fully effective defense response against fire blight upon ASM treatment was compromised, confirming the role of MdAGGs in fire blight resistance. Next, we coupled the pPPO16 promoter, endogenous to apple and known to be rapidly induced during E. amylovora infection, with the coding sequence of MdAGG10 to create apple lines with fire blight-inducible MdAGG10 expression. Early MdAGG10 expression in these lines significantly improved resistance to fire blight, and an additional ASM treatment further enhanced this resistance. In summary, we conclude that MdAGGs act as defense genes whose timely expression can provide effective resistance against E. amylovora.

plant biology↗

bioRxiv2023

Overexpression of an apple broad range agglutinating lectin does not promote in planta resistance to fire blight and bacterial wilt.

Lectins, a large group of proteins present in all kingdoms of life can bind reversibly to glycans. The roles of plant lectins are diverse and include resistance to biotic or abiotic stress, notably bacterial resistance. A gene family encoding amaranthin-like lectins termed MdAGGs in apple (Malus domestica) has been identified to be overexpressed upon treatment with the plant resistance inducer acibenzolar-S-methyl (ASM) which promotes enhanced resistance to the fire blight disease caused by Erwinia amylovora (Ea). In this study, we first screened the ability of purified MdAGG10 to agglutinate bacterial cells in vitro among a range of bacterial species. Several bacterial species, either Gram positive or negative, either plant- or human-pathogens were found to be agglutinated by MdAGG10 in acidic conditions. Apple and Arabidopsis lines constitutively overexpressing MdAGG10 were generated and evaluated for their resistance to, respectively, Ea and Ralstonia solanacearum, both plant pathogens that were found in our screening. Despite MdAGG10 protein accumulated in tissues of both apple and Arabidopsis lines, they remained susceptible to their respective pathogens. Interestingly, in vitro agglutination of Ea by MdAGG10 did not impair bacterial growth, suggesting that other plant molecules are involved in the resistance to fire blight triggered after an ASM treatment.

plant biology↗

bioRxiv2022

CRTC-1 Balances Histone Trimethylation and Acetylation to Promote Longevity

Loss of function during ageing is accompanied by transcriptional drift, altering gene expression and contributing to a variety of age-related diseases. CREB-regulated transcriptional coactivators (CRTCs) have emerged as key regulators of gene expression that might be targeted to promote longevity. Here, we define the role of the Caenorhabditis elegans CRTC-1 in the epigenetic regulation of longevity. Endogenous CRTC-1 binds chromatin factors, including components of the COMPASS complex, which trimethylates lysine 4 on histone H3 (H3K4me3). CRISPR editing of endogenous CRTC-1 reveals that the CREB-binding domain in neurons is specifically required for H3K4me3-dependent longevity. However, this effect is independent of CREB but instead acts via the transcription factor AP-1. Strikingly, CRTC-1 also mediates global histone acetylation levels, and this acetylation is essential for H3K4me3-dependent longevity. Indeed, overexpression of an acetyltransferase enzyme is sufficient to promote longevity in wild-type worms. CRTCs, therefore, link energetics to longevity by critically fine-tuning histone acetylation and methylation to promote healthy ageing.

genetics↗

bioRxiv2021

A Single-copy Knock In Translating Ribosome ImmunoPrecipitation (SKI TRIP) tool kit for tissue specific profiling of actively translated mRNAs in C. elegans.

Translating Ribosome Affinity Purification (TRAP) methods have emerged as a powerful approach to profile actively translated transcripts in specific cell and tissue types. Epitope tagged ribosomal subunits are expressed in defined cell populations and used to pull down ribosomes and their associated mRNAs, providing a snapshot of cell type-specific translation occurring in that space and time. Current TRAP toolkits available to the C. elegans community have been built using multi-copy arrays, randomly integrated in the genome. Here we introduce a Single-copy Knock In Translating Ribosome ImmunoPrecipitation (SKI TRIP) tool kit, a collection of C. elegans strains engineered by CRISPR in which tissue specific expression of FLAG tagged ribosomal subunit protein RPL-22 is driven by cassettes present in single copy from defined sites in the genome. In depth characterization of the SKI TRIP strains and methodology shows that 3xFLAG tagged RPL-22 expressed from its endogenous locus or within defined cell types incorporates into actively translating ribosomes and can be used to efficiently and cleanly pull-down cell type specific transcripts without impacting overall mRNA translation or fitness of the animal. We propose SKI TRIP use for the study of processes that are acutely sensitive to changes in translation, such as aging.

genetics↗

bioRxiv2021

Efficacy of longevity interventions in C. elegans is determined by early life activity of RNA splicing factors

Geroscience aims to target the aging process to extend healthspan. However, even isogenic individuals show heterogeneity in natural aging rate and responsiveness to pro-longevity interventions, limiting translational potential. Using in vivo mini gene reporters in isogenic C. elegans, we show that alternative splicing of mRNAs related to lipid metabolism in young animals is coupled to subsequent life expectancy. Further, activity of RNA splicing factors REPO-1 and SFA-1 early in life modulates effectiveness of specific longevity interventions via POD-2/ACC1 and regulation of lipid utilization. In addition, early inhibition of REPO-1 renders animals refractory to late onset suppression of the TORC1 pathway. Together these data suggest that activity of RNA splicing factors and the metabolic landscape early in life can modulate responsiveness to longevity interventions and may explain variance in efficacy between individuals. One Sentence SummaryEfficacy of pro-longevity interventions in C. elegans is determined by the activity of splicing factors and the lipid metabolic landscape early in the life of the individual.

genetics↗

bioRxiv2018

Single-copy Knock-In Loci for Defined Gene Expression in C. elegans

We have generated a single-copy knock-in loci for defined gene expression (SKI LODGE) system to insert any cDNA by CRISPR/Cas9 at defined safe harbors in the Caenorhabditis elegans genome. Utilizing a single crRNA guide, which also acts as a Co-CRISPR enrichment marker, any cDNA sequence can be introduced as a single integrated copy, regulated by different tissue-specific promoters. The SKI LODGE system provides a fast, economical and effective approach for generating single-copy non-silenced transgenes in C. elegans.

genetics↗