Abstract Bacterial type VI secretion systems (T6SSs) inject toxic effectors into adjacent eukaryotic and prokaryotic cells. It is generally thought that this process requires physical contact between the two Here, we provide evidence of contact-independent killing by a T6SS-secreted effector. We show pathogen Yersinia pseudotuberculosis uses T6SS (T6SS-3) to secrete nuclease effector kills other bacteria in vitro facilitates gut colonization mice. The (Tce1) small protein acts as Ca 2+ - Mg...

Abstract In flowering plants, pollen formation depends on the differentiation and interaction of two cell types in anther: reproductive cells, called microsporocytes, somatic cells that form tapetum. Previously, we cloned a specific gene, zm401 , from cDNA library generated mature Zea mays . Expression partial maize ectopic expression tobacco suggested it may play role anther development. Here present functional characterization this gene maize. Zm401 is expressed primarily anthers (tapetal...

Molecular Beacons (MBs) composed of 2′-O-methyl RNA (2Me) and phosphorothioate (PS) linkages throughout the backbone (2Me/PSFULL MBs) have enabled long-term imaging in living cells, but excess PS modification can induce nonspecific binding, causing false-positive signals. In this study, we evaluate intracellular stability MBs 2Me with various modifications, found that signals could be reduced to marginal levels when possess a fully PS-modified loop domain phosphodiester stem (2Me/PSLOOP MB)....

Analysis of RNA dynamics and localization at the single-molecule level in living cells has been predominantly achieved by engineering target RNAs with large insertions tandem repeat sequences that are bound protein-based or oligonucleotide-based fluorescent probes. Thus, individual tagged multiple probes, making them detectable fluorescence microscopy. Since may affect processes including trafficking localization, here we present a strategy to visualize single transcripts using molecular...

ABSTRACT Urease converts urea into ammonia and carbon dioxide, providing a nitrogen source for microbial growth serving as an important mechanism human bacterial pathogens to survive in acidic conditions, which can be regulated by many factors. As global regulator, the ferric uptake regulator (Fur) regulates series of genes pathways involved different cellular processes virulence enteric bacterium Yersinia pseudotuberculosis ( Yptb ). However, whether Fur urease activity was still unknown....

The evolutionarily conserved G-quadruplexes (G4s) are faithfully inherited and serve a variety of cellular functions such as telomere maintenance, gene regulation, DNA replication initiation, epigenetic regulation. Different from the Watson-Crick base-pairing found in duplex DNA, G4s formed via Hoogsteen base pairing very stable compact structures. Failure untangling them cell impedes DNA-based transactions leads to genome instability. Cells have evolved highly specific helicases resolve G4...

The clustered regularly interspersed short palindromic repeat (CRISPR) gene-editing system has been repurposed for live-cell genomic imaging, but existing approaches rely on fluorescent protein reporters, making sensitive and continuous imaging difficult. Here, we present a fluorophore-based that consists of nuclease-deactivated mutant the Cas9 (dCas9), molecular beacon (MB), an engineered single-guide RNA (sgRNA) harboring unique MB target sequence (sgRNA-MTS), termed CRISPR/MB....

Burkholderia thailandensis is a model organism for human pathogens mallei and pseudomallei. The study of B. peroxiredoxin helpful understanding the survival, pathogenic infection, antibiotic resistance its homologous species. Alkyl hydroperoxide reductase subunit C (AhpC) an important involved in oxidative damage defense. Here, we report that BthAhpC exhibits broad specificity peroxide substrates, including inorganic organic peroxides peroxynitrite. AhpC catalyzes reduction oxidants using...

Dental pain is a persistent, detrimental public health issue that requires better understanding of the mechanisms tooth and inflammation in order to develop more effective treatments. Calcitonin gene-related peptide (CGRP) dental pulp cells are promising candidates for mediating generating reparative tissues, respectively, but their behavior context pulpitis remains elusive. The mouse incisor Sonic hedgehog (Shh) secreted from sensory nerves continuously regenerate. However, it unknown...

Zur (zinc uptake regulator) is a significant member of the Fur (ferric superfamily, which widely distributed in bacteria. plays crucial roles zinc homeostasis and influences cell development environmental adaptation various species. Yersinia pseudotuberculosis Gram-negative enteric that pathogen usually serves as model organism pathogenicity studies. The regulatory effects on transporter ZnuABC protein secretion system T6SS have been documented Y. pseudotuberculosis. In this study,...

Abstract Purpose Based on the quantitative and qualitative features of CT imaging, a model for predicting invasiveness ground-glass nodules (GGNs) was constructed, which could provide reference value preoperative planning GGN patients. Materials methods Altogether, 702 patients with GGNs (including 748 GGNs) were included in this study. The operated between September 2020 July 2022 classified into training group ( n = 555), those August November validation 193). Clinical data imaging...

ScPif1 DNA helicase is the prototypical member of a 5′-to-3′ superfamily conserved from bacteria to human and plays various roles in maintenance genomic homeostasis. While many studies have been performed with eukaryotic Pif1 helicases, including yeast proteins, potential functions biochemical properties prokaryotic helicases remain largely unknown. Here, we report expression, purification analysis Bacteroides sp. 3_1_23 (BsPif1). BsPif1 binds large panel substrates and, particular,...

Replication protein A (RPA) is the major eukaryotic ssDNA-binding and has essential roles in genome maintenance. RPA binds to ssDNA through multiple modes, recent studies have suggested that RPA–ssDNA interaction dynamic. However, how alternates between different binding modes modifies structures this dynamic remains unknown. Here, we used single-molecule FRET systematically investigate human ssDNA. We show can adopt types of complexes with ssDNAs lengths, leading straightening or bending...

Significance Single-molecule localization microscopy (SMLM) is useful for deciphering dynamic organizations of structures densely labeled by specific proteins in the cellular context with nanoscopic resolution not attainable conventional imaging tools. Here we employed SMLM to investigate mechanism which HIV-1 viral RNA (vRNA) mediates assembly thousands Gag into a virus particle at plasma membrane. In contrast general notion that vRNA only triggers and dispensable subsequent assembly, found...

HIV-1 Gag proteins can multimerize upon the viral genomic RNA or multiple random cellular messenger RNAs to form a virus particle virus-like particle, respectively. To date, whether two types of particles via same multimerization process has remained unclarified. Using photoactivated localization microscopy illuminate organizations and dynamics at nanoscale, here, we showed that mediates in more cluster-centric, cooperative, spatiotemporally coordinated fashion, with ability drive dense...

The type VI secretion system (T6SS) is a widespread protein apparatus deployed by many Gram-negative bacterial species to interact with competitor bacteria, host organisms, and the environment. Yersinia pseudotuberculosis T6SS4 was recently reported be involved in manganese acquisition; however, underlying regulatory mechanism still remains unclear. In this study, we discovered that regulated ferric uptake regulator (Fur) response ions (Mn

As a key bacterial second messenger, cyclic di-GMP (c-di-GMP) regulates various physiological processes, such as motility, biofilm formation, and virulence. Cellular c-di-GMP levels are regulated by the opposing activities of diguanylate cyclases (DGCs) phosphodiesterases (PDEs). Beyond that, enzymatic metabolizing proteins controlled variety extracellular signals intracellular conditions. Here, we report that pdcA (BTH_II2363), pdcB (BTH_II2364), pdcC (BTH_II2365) cotranscribed in same...

HpaR, a MarR family transcriptional regulator, was first identified in Escherichia coli W for its regulation of the hpa-meta operon. Little else is known regarding functionality. Here, we report that Yersinia pseudotuberculosis, HpaR negatively regulates operon similar to W. To investigate additional functions RNA sequencing performed both wild-type and ΔhpaR mutant, which revealed type VI secretion system (T6SS) positively regulated by HpaR. T6SS4 important bacteria resisting environmental...

Microbial species often occur in complex communities and exhibit intricate synergistic antagonistic interactions. To avoid predation compete for favorable niches, bacteria have evolved specialized protein secretion systems. The type VI system (T6SS) is a versatile widely distributed among Gram-negative that translocates effectors into target cells or the extracellular milieu via various physiological processes. Pseudomonas aeruginosa an opportunistic pathogen responsible many diseases, it...