Biology subjects

Schulz, L. M.

Publications and source records attributed to Schulz, L. M..

5 recordsLinked to original sources

bioRxiv2025

Cysteine-rich receptor-like kinases mediate Wall Teichoic Acid perception in Arabidopsis

Plants and animals detect microbe-associated molecular patterns (MAMPs) to initiate defense responses. Both lineages employ pattern recognition receptors (PRRs), yet plant mechanisms for sensing Gram-positive bacterial MAMPs remain poorly understood. Wall teichoic acids (WTAs), anionic glycopolymers unique to Gram-positive bacteria, are shown here to activate salicylic acid signaling, defense priming, and a non-canonical programmed cell death in Arabidopsis. We demonstrate that glycosylated WTAs from diverse Gram-positive species elicit these responses and provide genetic evidence that their recognition depends on two members of the plant-specific CYSTEINE-RICH RECEPTOR-LIKE KINASE (CRK) gene family, which is absent in animals and whose function remains elusive. These findings reveal WTAs as a novel class of MAMPs in plants and highlight CRKs as key components in their perception.

plant biology↗

bioRxiv2023

Adaptation mechanisms of Listeria monocytogenes to quaternary ammonium compounds

Listeria monocytogenes is ubiquitously found in nature and can easily enter food-processing facilities due to contaminations of raw materials. Several countermeasures are used to combat contamination of food products, for instance the use of disinfectants that contain quaternary ammonium compounds, such as benzalkonium chloride (BAC) and cetyltrimethylammonium bromide (CTAB). In this study, we assessed the potential of the commonly used wildtype strain EGD-e to adapt to BAC and CTAB under laboratory growth conditions. All BAC-tolerant suppressors exclusively carried mutations in fepR or its promoter region likely resulting in the overproduction of the efflux pump FepA. In contrast, CTAB- tolerance was associated with mutations in sugR, which regulates the expression of the efflux pumps SugE1 and SugE2. L. monocytogenes strains lacking either FepA or SugE1/2 could still acquire tolerance towards BAC and CTAB. Genomic analysis revealed that the overproduction of the remaining efflux system could compensate for the deleted one. Even in the absence of both efflux systems, tolerant strains could be isolated, which all carried mutations in the diacylglycerol kinase encoding gene lmo1753 (dgkB). DgkB converts diacylglycerol to phosphatidic acid, which is subsequently re-used for the synthesis of phospholipids suggesting that alterations in membrane composition could be the third adaptation mechanism. Originality-Significance StatementSurvival and proliferation of Listeria monocytogenes in the food industry is an ongoing concern, and while there are various countermeasures to combat contamination of food products, the pathogen still successfully manages to withstand the harsh conditions present in food-processing facilities, resulting in reoccurring outbreaks, subsequent infection, and disease. To counteract the spread of L. monocytogenes it is crucial to understand and elucidate the underlying mechanism that permit their successful evasion. We here present various adaptation mechanisms of L. monocytogenes to withstand two important quaternary ammonium compounds.

microbiology↗

bioRxiv2023

Physiological studies on pH and salt tolerance and the utilization of diverse carbon sources by Listeria monocytogenes

The foodborne pathogen Listeria monocytogenes can grow in a wide range of environmental conditions. For the study of the physiology of this organism, several chemically defined media have been developed over the past decades. Here, we examined the ability of L. monocytogenes wildtype strains EGD-e and 10403S to grow under salt and pH stress in Listeria synthetic medium (LSM). Furthermore, we determined that a wide range of carbon sources could support growth of both wildtype strains in LSM. However, for hexose phosphate sugars such as glucose-1-phosphate, both L. monocytogenes strains need to be pre-grown under conditions, where the major virulence regulator PrfA is active. In addition, growth of both L. monocytogenes strains was observed when LSM was supplemented with the amino acid sugar N-acetylmannosamine (ManNAc). We were able to show that some of the proteins encoded in the operon lmo2795-nanE, such as the ManNAc-6-phosphate epimerase NanE, are required for growth in presence of ManNAc. The first gene of the operon, lmo2795, encodes a transcriptional regulator of the RpiR family. Using electrophoretic mobility shift assays and quantitative real time PCR analysis, we were able to show that Lmo2795 binds to the promoter region of the operon lmo2795-nanE and activates its expression. Originality-Significance StatementCurrent knowledge of growth and survival of the human pathogen Listeria monocytogenes under diverse stress conditions is mostly generated in complex medium, a condition that is rarely found in the environment or host of the pathogen. Our work contributes to the characterization of the physiology of L. monocytogenes grown under nutrient limiting conditions and its growth requirements with regards to metabolizable carbon sources.

microbiology↗

bioRxiv2022

Imbalance of peptidoglycan biosynthesis alters the cell surface charge of Listeria monocytogenes

The bacterial cell wall is composed of a thick layer of peptidoglycan and cell wall polymers, which are either embedded in the membrane or linked to the peptidoglycan backbone and referred to as lipoteichoic acid (LTA) and wall teichoic acid (WTA), respectively. Modifications of the peptidoglycan or WTA backbone can alter the susceptibility of the bacterial cell towards cationic antimicrobials and lysozyme. The human pathogen Listeria monocytogenes is intrinsically resistant towards lysozyme, mainly due to deacetylation and O-acetylation of the peptidoglycan backbone via PgdA and OatA. Recent studies identified additional factors, which contribute to the lysozyme resistance of this pathogen. One of these is the predicted ABC transporter, EslABC. An eslB mutant is hyper-sensitive towards lysozyme, likely due to the production of thinner and less O-acetylated peptidoglycan. Using a suppressor screen, we show here that suppression of eslB phenotypes could be achieved by enhancing peptidoglycan biosynthesis, reducing peptidoglycan hydrolysis or alterations in WTA biosynthesis and modification. The lack of EslB also leads to a higher negative surface charge, which likely stimulates the activity of peptidoglycan hydrolases and lysozyme. Based on our results, we hypothesize that the portion of cell surface exposed WTA is increased in the eslB mutant due to the thinner peptidoglycan layer and that latter one could be caused by an impairment in UDP-N-acetylglucosamine (UDP-GlcNAc) production or distribution.

microbiology↗

bioRxiv2020

The YtrBCDEF ABC transporter is involved in the control of social activities in Bacillus subtilis

Bacillus subtilis develops genetic competence for the uptake of foreign DNA when cells enter the stationary phase and a high cell density is reached. These signals are integrated by the competence transcription factor ComK which is subject to transcriptional, post-transcriptional and post-translational regulation. Many proteins are involved in the development of competence, both to control ComK activity and to mediate DNA uptake. However, the precise function they play in competence development is often unknown. In this study, we have tested whether proteins required for genetic transformation play a role in the activation of ComK or rather downstream of competence gene expression. While these possibilities could be distinguished for most of the tested factors, two proteins (PNPase and the transcription factor YtrA) are required both for full ComK activity and for the downstream processes of DNA uptake and integration. Further analyses of the role of the transcription factor YtrA for the competence development revealed that the constitutive expression of the YtrBCDEF ABC transporter in the ytrA mutant causes the loss of genetic competence. Moreover, constitutive expression of this ABC transporter also interferes with biofilm formation. Since the ytrGABCDEF operon is induced by cell wall-targeting antibiotics, we tested the cell wall properties upon overexpression of the ABC transporter and observed an increased thickness of the cell wall. The composition and properties of the cell wall are important for competence development and biofilm formation, suggesting that the increased cell wall thickness as a result of YtrBCDEF overexpression causes the observed phenotypes.

microbiology↗