Michael Wrzaczek

The Czech Academy of Sciences | AVCR · Biology Centre

Cysteine-rich receptor-like kinases (CRKs) are transmembrane proteins characterized by the presence of two domains of unknown function 26 (DUF26) in their ectodomain. The CRKs form one of the largest groups of receptor-like protein kinases in plants, but their biological functions have so far remained largely uncharacterized. We conducted a large-s...

In plants, receptor-like kinases (RLKs) and extracellular reactive oxygen species (ROS) contribute to the communication between the environment and the interior of the cell. Apoplastic ROS production is a frequent result of RLK signaling in a multitude of cellular processes. However, it is as yet unclear how ROS signaling downstream of receptor act...

Large protein families are a prominent feature of plant genomes and their size variation is a key element for adaptation. However, gene and genome duplications pose difficulties for functional characterization and translational research. Here we infer the evolutionary history of the DOMAIN OF UNKNOWN FUNCTION (DUF) 26-containing proteins. The DUF26...

Reactive oxygen species (ROS) are important messengers in eukaryotic organisms and their production is tightly controlled. Active extracellular ROS production by NADPH oxidases in plants is triggered by receptor-like protein kinase (RLK)-dependent signaling networks. Here we show that the cysteine-rich RLK CRK2 kinase activity is required for plant...

Reactive oxygen species (ROS) are essential for life and are involved in the regulation of almost all biological processes. ROS production is critical for plant development, response to abiotic stresses and immune responses. Here, we focus on recent discoveries in ROS biology emphasizing abiotic and biotic stress responses. Recent advancements have...

Reactive oxygen species (ROS) produced by plant NADPH oxidases, respiratory burst oxidase homologs (RBOHs), play key roles in biotic and abiotic stress responses and development in plants. While properly controlled amounts of ROS function as signaling molecules, excessive accumulation of ROS can cause undesirable side effects due to their ability t...

Continuous reprograming of gene expression in response to environmental signals in plants is achieved through signaling hub proteins that integrate external stimuli and transcriptional responses. RADICAL-INDUCED CELL DEATH1 (RCD1) functions as a nuclear hub protein, which interacts with a variety of transcription factors with its C-terminal RST dom...

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Perception of biotic and abiotic stresses often leads to stomatal closure in plants1,2. Rapid influx of calcium ions (Ca²⁺) across the plasma membrane plays an important role in this response, but the identity of Ca²⁺ channels involved has remained elusive3,4. Here, we report that the Arabidopsis thaliana Ca²⁺-permeable channel OSCA1.3 controls sto...

The multinational Arabidopsis research community is highly collaborative and over the past thirty years these activities have been documented by the Multinational Arabidopsis Steering Committee (MASC). Here, we (a) highlight recent research advances made with the reference plant Arabidopsis thaliana ; (b) provide summaries from recent reports submi...

Programmed cell death regulates developmental and stress responses in eukaryotes. Golgi anti-apoptotic proteins (GAAPs) are evolutionarily conserved cell death regulators. Human and viral GAAPs inhibit apoptosis and modulate intracellular Ca ²⁺ fluxes, and viral GAAPs form cation-selective channels. Although most mammalian cell death regulators are...

In the version of this article initially published, there was a mistake in the calculation of the nucleotide mutation rate per site per generation: 1 × 10⁻⁹ mutations per site per generation was used, whereas 9.5 × 10⁻⁹ was correct. This error affects the interpretation of population-size changes over time and their possible correspondence with kno...

High salinity is an increasingly prevalent source of stress to which plants must adapt. The receptor-like protein kinases (RLKs), including members of the cysteine-rich receptor-like kinase (CRK) subfamily, are a highly expanded family of transmembrane proteins in plants that are largely responsible for communication between cells and the extracell...

Reactive oxygen species (ROS) are important messengers in eukaryotic organisms and their production is tightly controlled. Active extracellular ROS production by NADPH oxidases in plants is triggered by receptor-like protein kinase (RLK)-dependent signaling networks. Here we show that the cysteine-rich RLK CRK2 kinase activity is required for plant...

High salinity has become an increasingly prevalent source of stress to which plants need to adapt. The receptor-like protein kinases (RLKs), including the cysteine-rich receptor-like kinase (CRK) subfamily, are a highly expanded family of transmembrane proteins in plants and are largely responsible for communication between cells and the extracellu...

Reactive oxygen species (ROS)-dependent signaling pathways from chloroplasts and mitochondria merge at the nuclear protein RADICAL-INDUCED CELL DEATH1 (RCD1). RCD1 interacts in vivo and suppresses the activity of the transcription factors ANAC013 and ANAC017, which mediate a ROS-related retrograde signal originating from mitochondrial complex III....

During plant vascular development, xylem tracheary elements (TEs) form water-conducting, empty pipes through genetically regulated cell death. Cell death is prevented from spreading to non-TEs by unidentified intercellular mechanisms, downstream of METACASPASE9 (MC9)-mediated regulation of autophagy in TEs. Here, we identified differentially abunda...

The use of draft genomes of different species and re-sequencing of accessions and populations are now a common tool for plant biology research. The de novo assembled draft genomes make it possible to identify pivotal divergence points in the plant lineage and provide an opportunity to investigate the genomic basis and timing of biological innovatio...

Large protein families are a prominent feature of plant genomes and their size variation is a key element for adaptation in plants. Here we infer the evolutionary history of a representative protein family, the DOMAIN OF UNKNOWN FUNCTION (DUF) 26-containing proteins. The DUF26 first appeared in secreted proteins. Domain duplications and rearrangeme...

Table S6 Expression values of Arabidopsis thaliana SA and JA marker genes shown in Fig. 4 and detailed analysis of differentially expressed genes in Pro 35S :HS:HaRxL106 line #2 and rcd1‐1 using GO term annotation and analysis of cis‐regulatory elements

Fig. S1 Transcriptome profiling of Arabidopsis thaliana Pro 35S :HS:HaRxL106 line #2 and knock‐out mutants of the Hyaloperonospora arabidopsidis HaRxL106‐interacting proteins MOS6, ASIL1 and RCD1. Fig. S2 Sequence coverage of Nicotiana benthamiana RCD1, Arabidopsis thaliana RCD1 and A. thaliana SRO1 identified by peptide fingerprinting in immunopr...

Table S1 Quantification of Hyaloperonospora arabidopsidis Noco2 conidiophores on adult leaves of Arabidopsis thaliana and statistical analysis based on two independent biological experiments Table S2 Quantification of Arabidopsis thaliana seedling hypocotyl length and statistical analysis based on three independent biological experiments Table S3...

Table S11 X‐ray data collection, refinement, and validation statistics for the Arabidopsis thaliana RCD1 PARP domain structure

Table S15 Phosphopeptides identified in immunoprecipitates of the Arabidopsis thaliana RCD1 WWE‐linker bait protein expressed in Nicotiana benthamiana in three independent biological experiments

Methods S1 Methods for Arabidopsis thaliana Paraquat treatment and SA quantification, nucleic acid extraction and quantification, protein extraction and purification, microscopy, protein crystallography and protein mass spectrometry.

Table S12 Effect of different 6(5H)‐phenanthridinone concentrations on the thermal stability of the Arabidopsis thaliana RCD1 and Homo sapiens PARP1 PARP domains as determined by thermal shift assays and monitored in three independent experiments

Table S14 List of proteins identified in immunoprecipitates of the Arabidopsis thaliana RCD1 WWE‐linker bait protein expressed in Nicotiana benthamiana

The oomycete pathogen Hyaloperonospora arabidopsidis (Hpa) causes downy mildew disease on Arabidopsis. To colonize its host, Hpa translocates effector proteins that suppress plant immunity into infected host cells. Here, we investigate the relevance of the interaction between one of these effectors, HaRxL106, and Arabidopsis RADICAL‐INDUCED CELL DE...

Signaling from chloroplasts and mitochondria, both dependent on reactive oxygen species (ROS), merge at the nuclear protein RADICAL-INDUCED CELL DEATH1 (RCD1). ROS produced in the chloroplasts affect the abundance, thiol redox state and oligomerization of RCD1. RCD1 directly interacts in vivo with ANAC013 and ANAC017 transcription factors, which ar...

Small signalling peptides have emerged as important cell to cell messengers in plant development and stress responses. However, only a few of the predicted peptides have been functionally characterized. Here, we present functional characterization of two members of the IDA-LIKE (IDL) peptide family in Arabidopsis thaliana, IDL6 and IDL7. Localizati...

Analysis of gene families and identification of homologous genes are important for phylogenetic analysis and for translating results from model to crop species. While numerous plant genomes have been sequenced and made available, the identification of gene models can be difficult, in particular for large gene families arranged in tandem repeats or...

Silver birch (Betula pendula) is a pioneer boreal tree that can be induced to flower within 1 year. Its rapid life cycle, small (440-Mb) genome, and advanced germplasm resources make birch an attractive model for forest biotechnology. We assembled and chromosomally anchored the nuclear genome of an inbred B. pendula individual. Gene duplicates from...

The oomycete pathogen Hyaloperonospora arabidopsidis (Hpa) causes downy mildew disease on Arabidopsis. During infection, Hpa like other biotrophic pathogens, suppresses activation of plant innate immunity by translocating effector proteins into host cells. Some of these effectors localize to the host cell nucleus where they may manipulate transcrip...

A powerful method to study gene function is expression or overexpression in an inducible, cell type-specific system followed by observation of consequent phenotypic changes and visualization of linked reporters in the target tissue. Multiple inducible gene overexpression systems have been developed for plants, but very few of these combine plant se...

Cysteine-rich receptor-like kinases (CRKs) are transmembrane proteins characterized by the presence of two domains of unknown function 26 (DUF26) in their ectodomain. The CRKs form one of the largest groups of receptor-like protein kinases in plants, but their biological functions have so far remained largely uncharacterized. We conducted a large-s...

With the tremendous progress of the past decades, molecular plant science is becoming more unified than ever. We now have the exciting opportunity to further connect subdisciplines and understand plants as whole organisms, as will be required to efficiently utilize them in natural and agricultural systems to meet human needs. The subfields of photo...

Recognition of extracellular peptides by plasma membrane-localized receptor proteins is commonly used in signal transduction. In plants, very little is known about how extracellular peptides are processed and activated in order to allow recognition by receptors. Here, we show that induction of cell death in planta by a secreted plant protein GRIM R...

Significance: Reactive oxygen species (ROS), important signaling molecules in plants, are involved in developmental control and stress adaptation. ROS production can trigger broad transcriptional changes; however, it is not clear how specificity in transcriptional regulation is achieved. Recent advances: A large collection of public transcriptom...

One of the largest groups of recept or-like protein kinases (RLKs) are the CYSTEINE-RICH RLKs (CRKs) with 44 members in Arabidopsis. CRK expression is regulated in response to variou s stresses that lead to specific alter- ations in ROS metabolism (Wrzaczek et al., 2010). CRKs are characterized by conserved cy steine motifs in the ecto- domain pote...

The production of reactive oxygen species (ROS) in different plant subcellular compartments is the hallmark of the response to many stress stimuli and developmental cues. The past two decades have seen a transition from regarding ROS as exclusively cytotoxic agents to being considered as reactive compounds which participate in elaborate signaling n...

Transcriptional regulation of gene expression is one major determinant of developmental control and stress adaptation in virtually all living organisms. In recent years numerous transcription factors controlling various aspects of plant life have been identified. The activity of transcription factors needs to be regulated to prevent unspecific, pro...

Light is an important environmental factor that modulates acclimation strategies and defense responses in plants. We explored the functional role of the regulatory subunit B'γ (B'γ) of protein phosphatase 2A (PP2A) in light-dependent stress responses of Arabidopsis (Arabidopsis thaliana). The predominant form of PP2A consists of catalytic subunit C...

Lower and upper percentiles and p-values. The raw normalized cycle differences (ΔΔCt) for all experiments, their average, standard deviation, geometric mean, lower and upper percentile and the Benjamini-Hochberg False Discovery Rate-corrected p-value for all experiments is shown in the Excel File. Each Excel worksheet represents data for a Figure s...

Transcriptional regulation of the CRKs in response to flg22. 14-day old Arabidopsis Col-0 were treated with 100 nM flg22 and samples taken after 30 and 60 minutes (water-treated control samples have been harvested at the same time points in parallel). Expression of several CRKs was analyzed by qPCR. Transcript levels were calculated by comparison w...

List of CRKs for promoter motifs in table 2. This file lists the AGI codes for the CRKs containing the promoter motif combinations shown in table 2.

Primer sequences for qPCR analysis. All primer sequences used for qPCR analysis in the manuscript plus the experimentally determined primer amplification efficiencies Ex are listed.

Plant Receptor-like/Pelle kinases (RLK) are a group of conserved signalling components that regulate developmental programs and responses to biotic and abiotic stresses. One of the largest RLK groups is formed by the Domain of Unknown Function 26 (DUF26) RLKs, also called Cysteine-rich Receptor-like Kinases (CRKs), which have been suggested to play...

Primers used for analysis. All primer sequences used for qPCR anaylsis in the manuscript are listed in this file.

Active site alignments for all plant SROs. Alignments of the region around the active site catalytic triad of all SROs analyzed here. Alignments were hand-adjusted according to the positions of conserved folds in AtRCD1, AtPARP1 and AtPARP2 as in figure 10, only the regions immediately surrounding the predicted catalytic amino acids (highlighted in...