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The mechanisms involved in the virulence of Yersinia pestis, the plague pathogen, are not fully understood. In previous research, we found that a Y. pestis mutant lacking the HicB3 (YPO3369) putative orphan antitoxin was attenuated for virulence in a murine model of bubonic plague. Toxin-antitoxin systems (TASs) are widespread in prokaryotes. Most bacterial species possess many TASs of several types. In type II TASs, the toxin protein is bound and neutralized by its cognate antitoxin protein in the cytoplasm. Here we identify the hicA3 gene encoding the toxin neutralized by HicB3 and show that HicA3-HicB3 constitutes a new functional type II TAS in Y. pestis. Using biochemical and mutagenesis-based approaches, we demonstrate that the HicA3 toxin is an RNase with a catalytic histidine residue. HicB3 has two functions: it sequesters and neutralizes HicA3 by blocking its active site, and it represses transcription of the hicA3B3 operon. Gel shift assays and reporter fusion experiments indicate that the HicB3 antitoxin binds to two operators in the hicA3B3 promoter region. We solved the X-ray structures of HicB3 and the HicA3-HicB3 complex; thus, we present the first crystal structure of a TA complex from the HicAB family. HicB3 forms a tetramer that can bind two HicA3 toxin molecules. HicA3 is monomeric and folds as a double-stranded-RNA-binding domain. The HicB3 N-terminal domain occludes the HicA3 active site, whereas its C-terminal domain folds as a ribbon-helix-helix DNA-binding motif.
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http://dx.doi.org/10.1128/JB.01932-14 | DOI Listing |
Microbiol Spectr
December 2024
Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, Ohio, USA.
Automated continuous monitoring blood culture instruments identify metabolism byproducts and flag blood culture bottles as "positive." A Gram stain is used to visualize and characterize the microbial growth in the broth and initiate additional testing. When no organisms are seen (NOS) on Gram stain, in our laboratory, bottles are reevaluated with a Wayson stain, a rapid one-step stain that provides contrast between organisms and the background, especially in Gram-negative organisms.
View Article and Find Full Text PDFTwo live attenuated vaccines (LAVs), LMA and LMP, were evaluated alone or in combination with a trivalent adenoviral vector-based vaccine (Ad5-YFV) for their efficacy and immune responses in wild type (WT) and interferon gamma (IFNγ) knockout (KO) mice in a C57BL/6 background. While LMA and LMP are triple deletion mutants of CO92 strain, Ad5-YFV incorporates three protective plague immunogens. An impressive 80-100% protection was observed in all vaccinated animals against highly lethal intranasal challenge doses of parental CO92.
View Article and Find Full Text PDFMath Biosci Eng
October 2024
Department of Applied Mathematics, Hong Kong Polytechnic University, Hong Kong SAR, China.
Identifying epidemic-driving factors through epidemiological modeling is a crucial public health strategy that has substantial policy implications for control and prevention initiatives. In this study, we employ dynamic modeling to investigate the transmission dynamics of pneumonic plague epidemics in Hong Kong from 1902 to 1904. Through the integration of human, flea, and rodent populations, we analyze the long-term changing trends and identify the epidemic-driving factors that influence pneumonic plague outbreaks.
View Article and Find Full Text PDFbioRxiv
December 2024
Department of Molecular Genetics and Microbiology, School of Medicine, Duke University, Durham, NC, USA.
is the gram-negative bacterium responsible for plague, one of the deadliest and most feared diseases in human history. This bacterium is known to infect phagocytic cells, such as dendritic cells and macrophages, but interactions with non-phagocytic cells of the adaptive immune system are frequently overlooked despite the importance they likely hold for human infection. To discover human genetic determinants of infection, we utilized nearly a thousand genetically diverse lymphoblastoid cell lines in a cellular genome-wide association study method called Hi-HOST (High-throughput Human in-vitrO Susceptibility Testing).
View Article and Find Full Text PDFMicrobiol Resour Announc
December 2024
Department of Microbiology and Immunology, University of Louisville School of Medicine, Louisville, Kentucky, USA.
is the etiological agent of human plague. However, certain evolutionarily divergent subspecies have different host specificities and virulence capacity compared to the more commonly studied strains with pandemic potential. This resource examines 10 diverse isolates representing some of the most understudied subspecies commonly referred to as Pestoides.
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