A5004480637- Bacterial Genetics and Biotechnology
- RNA and protein synthesis mechanisms
- Bacteriophages and microbial interactions
- Mycobacterium research and diagnosis
- Endoplasmic Reticulum Stress and Disease
- Antimicrobial Peptides and Activities
- Cancer Research and Treatments
- CRISPR and Genetic Engineering
- Heat shock proteins research
- Enzyme Structure and Function
- RNA modifications and cancer
- Antibiotic Resistance in Bacteria
- Tuberculosis Research and Epidemiology
- Genomics and Phylogenetic Studies
- Protein Structure and Dynamics
- Advanced biosensing and bioanalysis techniques
- Bacterial biofilms and quorum sensing
- Animal Genetics and Reproduction
- Protist diversity and phylogeny
Czech Academy of Sciences
2014-2024
Czech Academy of Sciences, Institute of Microbiology
2014-2024
Charles University
2017
Czech Academy of Sciences, Institute of Biotechnology
2014
Czech Academy of Sciences, Institute of Macromolecular Chemistry
2014
Bacillus subtilis cells are well suited to study how bacteria sense and adapt proteotoxic stress such as heat, since temperature fluctuations a major challenge soil-dwelling bacteria. Here, we show that the alarmones (p)ppGpp, known second messengers of nutrient starvation, also involved in heat response development thermo-resistance. Upon heat-shock, intracellular levels (p)ppGpp rise rapid but transient manner. The heat-induced is primarily produced by ribosome-associated alarmone...
Bacterial RNA polymerase (RNAP) is an essential multisubunit protein complex required for gene expression. Here, we characterize YvgS (HelD) from Bacillus subtilis, a novel binding partner of RNAP. We show that HelD interacts with RNAP-core between the secondary channel RNAP and alpha subunits. Importantly, demonstrate stimulates transcription in ATP-dependent manner by enhancing transcriptional cycling elongation. stimulatory effect can be amplified small subunit RNAP, delta. In vivo, not...
The increase in the number of bacterial strains resistant to known antibiotics is alarming. In this study we report synthesis novel compounds termed Lipophosphonoxins II (LPPO II). We show that LPPO display excellent activities against Gram-positive and -negative bacteria, including pathogens multiresistant strains. describe their mechanism action-plasmatic membrane pore-forming activity selective for bacteria. Importantly, neither damage nor cross eukaryotic plasmatic at bactericidal...
Spx is a Bacillus subtilis transcription factor that interacts with the alpha subunits of RNA polymerase. It can activate thiol stress response regulon and interfere activation many developmental processes. Here, we show central player orchestrating heat shock by up-regulating relevant genes as revealed comparative transcriptomic experiments. Moreover, these experiments potential to inhibit translation-related genes. By in vivo vitro experiments, confirmed from rRNA. This inhibition depended...
The alarming rise of bacterial antibiotic resistance requires the development new compounds. Such compounds, lipophosphonoxins (LPPOs), were previously reported to be active against numerous species, but serum albumins abolished their activity. Here we describe synthesis and evaluation novel antibacterial compounds termed LEGO-LPPOs, loosely based on LPPOs, consisting a central linker module with two attached connector modules either side. are then decorated polar hydrophobic modules. We...
The expression of rRNA is one the most energetically demanding cellular processes and, as such, it must be stringently controlled. Here, we report that DNA topology, i.e., level supercoiling, plays a role in regulation Bacillus subtilis σA-dependent promoters growth phase-dependent manner. more negative supercoiling exponential phase stimulates transcription from promoters, and relaxation stationary contributes to cessation their activity. Novobiocin treatment B. cells relaxes decreases...
Mycobacterial HelD is a transcription factor that recycles stalled RNAP by dissociating it from nucleic acids and, if present, the antibiotic rifampicin. The rescued RNAP, however, must disengage to participate in subsequent rounds of transcription. mechanism release unknown. We show Mycobacterium smegmatis forms complex with associated primary sigma σ
The HelD is a helicase-like protein binding to Bacillus subtilis RNA polymerase (RNAP), stimulating transcription in an ATP-dependent manner. Here, our small angle X-ray scattering data bring the first insights into structure: compact shape and undergoes conformational change upon substrate analog binding. Furthermore, domain structure delineated, partial model of presented. In addition, unique N-terminal characterized as essential for its transcription-related function but not ATPase...
Sigma factors bind and direct the RNA polymerase core to specific promoter sequences, alternative sigma transcription of different regulons genes. Here, we study pBS32 plasmid-encoded factor SigN
Rifampicin is a clinically important antibiotic that binds to, and blocks the DNA/RNA channel of bacterial RNA polymerase (RNAP). Stalled, nonfunctional RNAPs can be removed from DNA by HelD proteins; this for maintenance genome integrity. Recently, it was reported proteins high G+C Actinobacteria, called HelR, are able to dissociate rifampicin-stalled provide rifampicin resistance. This achieved ability HelR RNAP. The HelR-mediated mechanism resistance discussed here, roles HelD/HelR in...
Abstract Here, B. subtilis was used as a model organism to investigate how cells respond and adapt proteotoxic stress conditions. Our experiments suggested that the stringent response, caused by raised levels of (p)ppGpp alarmone, plays role during thermotolerance development heat shock response. Accordingly, our revealed rapid increase cellular upon well salt- oxidative stress. Strains lacking exhibited increased sensitivity, while conferred tolerance heat- During development, response...
SUMMARY RNA synthesis is central to life, and polymerase depends on accessory factors for recovery from stalled states adaption environmental changes. Here we investigated the mechanism by which a helicase-like factor HelD recycles polymerase. We report cryo-EM structure of an unprecedented complex between Mycobacterium smegmatis HelD. The crescent-shaped simultaneously penetrates deep into two channels that are responsible DNA binding substrate delivery active site, thereby locking in...
Bacterial RNA polymerase (RNAP) is an essential multisubunit enzyme performing transcription.Regulation of this process secured through the stage-dependent interactions RNAP with different factors (mostly proteins).Here we report structurefunction analysis functional complexes between and a unique helicase-like factor HelD [1] which present in many Gram-positive bacteria (e.g.Bacillus subtilis Mycobacterium smegmatis) [2, 3].HelD forms tightly bound RNAP.It simultaneously penetrates into...