Biology subjects

Romeis, T.

Publications and source records attributed to Romeis, T..

5 recordsLinked to original sources

bioRxiv2024

The plant specific cohesin subunit SYN4 contributes to 3D genome organization and activity

O_LIChromatin architecture in the cells of animals and fungi influences gene expression. The molecular factors that influence higher genome architecture in plants and their effects on gene expression remain unknown. Cohesin complexes, conserved in eukaryotes, are essential factors in genome structuring. Here, we investigated the relevance of the plant-specific somatic cohesin subunit SYN4 for chromatin organisation in Arabidopsis thaliana. C_LIO_LIPlants mutated in SYN4 were studied using HRM, Hi-C, RNA sequencing, untargeted and targeted metabolomics and physiological assays to understand the role of this plant-specific cohesin. C_LIO_LIWe show that syn4 mutants exhibit altered intra- and interchromosomal interactions, expressed as sharply reduced contacts between telomeres and chromosome arms but not between centromeres, and differences in the placement and number of topologically associated domains (TADs)-like structures. By transcriptome sequencing, we also show that syn4 mutants have altered gene expression, including numerous genes that control abiotic stress responses. C_LIO_LIThe response to drought stress in Arabidopsis is strongly influenced by the genome structure in syn4 mutants, potentially due to altered expression of CYP707A3, an ABA 8-hydroxylase. C_LI In briefThe 3D architecture of the genome extensively influences gene expression in animals and fungi. We show that the plant-specific cohesin subunit SYN4 affects intra- and interchromosomal interactions including telomere clustering, with consequences for the expression of genes of transient and induced biological pathways and the biosynthesis of bioactive compounds. Highly condensed genome structures at the CYP707A3 locus positively affects the stress response to water deprivation regulated by abscisic acid.

genetics↗

bioRxiv2023

A bi-kinase module sensitizes and potentiates plant immune signaling

Systemic signaling is an essential hallmark of multicellular life. Pathogen encounter occurs locally but triggers organ-scale and organismic immune responses. In plants, elicitor perception provokes systemically expanding Ca2+ and H2O2 signals conferring immunity. Here, we identify a Ca2+ sensing bi-kinase module as becoming super-activated through mutual trans-phosphorylation and imposing synergistically enhanced NADPH oxidase activation. A combined two-layer bi-kinase/substrate phospho-code allows for sensitized signaling initiation already by near-resting elevations of Ca2+ concentration at the infection site. Subsequently, it facilitates further signal wave proliferation with minimal amplitude requirement, triggering protective defense responses throughout the plant. Our study reveals how plants build and perpetuate trans-cellular immune signal proliferation while avoiding disturbance of ongoing cellular signaling along the path of response dissemination. One sentence summaryMutual trans-activation of a Ca2+ sensing bi-kinase module potentiates NADPH oxidase activation to facilitate systemic immunity.

plant biology↗

bioRxiv2023

Imaging of plant calcium-sensor kinase conformation monitors real time calcium decoding in planta

Changes in cytosolic calcium concentration are among the earliest reactions to a multitude of stress cues. Whereas a plethora of calcium-permeable channels may generate distinct calcium signatures and contribute to response specificities, the mechanisms by which calcium signatures are decoded is poorly understood. Here we develop a genetically encoded, FRET-based reporter that visualizes the conformational change of calcium-dependent protein kinases (CDPKs/CPKs), preceding kinase activation, for calcium-dependent AtCPK21 and calcium-independent AtCPK23. In pollen tubes, naturally displaying a physiological calcium range, CPK21-FRET, but not CPK23-FRET, report activity oscillations with similar features to cytosolic calcium, suggesting an isoform-specific calcium dependency and reversibility of the conformational change. In guard cells CPK21-FRET identifies CPK21 as a decoder of signal-specific calcium signatures in response to ABA and flg22. Based on this data, CDPK-FRET stands as a novel approach for tackling real-time live-cell calcium decoding in a multitude of plant developmental and stress responses.

biochemistry↗

bioRxiv2019

Prioritization of abiotic and biotic stress responses by direct linkage of ABI1 phosphatase and CPK5 calcium-dependent protein kinase

In nature plants are constantly challenged by simultaneous abiotic and biotic stresses, and under conflicting stress scenarios prioritization of stress responses is required for plant survival. Calcium-dependent protein kinase CPK5 is a central hub in local and distal immune signaling, required for hormone salicylic acid (SA)-dependent immunity and pathogen resistance. Here we show that CPK5-dependent immune responses and pathogen resistance are inhibited upon abscisic acid (ABA) treatment or in genetic mutant backgrounds lacking PP2C phosphatase activities including abi1-2, whereas immune responses are enhanced by co-expression of active ABI1 phosphatase variants. Biochemical studies and mass spectrometry-based phospho-site analysis reveal a direct ABI1 phosphatase-catalyzed de-phosphorylation of CPK5 auto-phosphorylation site T98. Mimicking continuous de-phosphorylation in CPK5T98A leads to enhanced ROS production and more resistant plants, mimicking the auto-phosphorylated status in CPK5T98D, reduces CPK5-mediated immune responses. Mechanistic insight identifies differential phosphorylation at T98 in the N-terminal domain of CPK5 to control the level of interaction between the kinase and its substrate protein rather than CPK5 catalytic activity. Thus, CPK5-catalyzed immune signaling may become discontinued even at an elevated cytoplasmic calcium concentration. Our work reveals an elegant mechanism for stress response prioritization in plants: The ABA-dependent phosphatase ABI1, negative regulator of abiotic responses, functions as positive regulator of biotic stress responses, stabilizing CPK5-dependent immune signaling in the absence of ABA. Continuous pathogen survey activates plant immunity in environmentally friendly conditions, whereas under severe abiotic stress the phosphatase/kinase pair prohibits immune signaling through a direct biochemical switch involving two key regulatory enzymes of these antagonistic pathways. Significance StatementPlants challenged by simultaneous abiotic and biotic stresses must prioritize in conflicting scenarios to guarantee survival. Pathogen resistance and immune memory depends on the phytohormone salicylic acid (SA). Adaptation to abiotic stress signaling involves the phytohormone abscisic acid (ABA). We identify a direct biochemical switch by which ABA-mediated abiotic signaling prioritizes over SA-dependent immune responses via reversible phosphorylation at a single protein mark involving two key regulatory enzymes of these antagonistic pathways. Phosphatase ABI1 de-phosphorylates calcium-dependent protein kinase CPK5 at an auto-phosphorylation site T98, which effects the interaction efficiency between the kinase and its substrate. Under abiotic stress ABA mediates phosphatase inhibition, which facilitates prolonged auto-phosphorylation of CPK5, preventing CPK5 substrate interaction and ultimately stop CPK5-mediated immune signaling.

plant biology↗

bioRxiv2019

Calcium-dependent protein kinase 5 links calcium-signaling with SARD1-dependent immune memory in systemic acquired resistance

- Systemic acquired resistance (SAR) prepares infected plants for faster and stronger defense activation upon subsequent attacks. SAR requires an information relay from primary infection to distal tissue and the initiation and maintenance of a self-maintaining phytohormone salicylic acid (SA)-defense loop. - In spatial and temporal resolution we show that calcium-dependent protein kinase CPK5 contributes to immunity and SAR. In local basal resistance CPK5 functions upstream of SA-synthesis, -perception, and -signaling. In systemic tissue, enhanced CPK5 signaling leads to an accumulation of SAR marker genes including transcription factor Systemic Acquired Resistance Deficient 1 (SARD1). - Plants of enhanced CPK5-, but not CPK6-, signaling display a super-priming phenotype of enhanced resistance toward a secondary bacterial infection. In sard1 background, CPK5-mediated basal resistance is still mounted but systemic super-priming is lost. - The biochemical analysis determines CPK5 half maximal kinase activity for calcium K50 [Ca2+] to [~]100 nM close to the cytoplasmic resting level. This low activation threshold uniquely qualifies CPK5 to decode subtle changes in calcium prerequisite to immune signal relay and to onset and maintenance of priming at later time points in distal tissue. Our data explain why CPK5 functions as a hub in basal and systemic plant immunity.

plant biology↗