An expanded genetic toolkit for inducible expression and targeted gene silencing inRickettsia parkeri DOI Creative Commons
Jon McGinn,

A. Wen,

Desmond L. Edwards

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 15, 2024

ABSTRACT Pathogenic species within the Rickettsia genus are transmitted to humans through arthropod vectors and cause a spectrum of diseases ranging from mild life-threatening. Despite rickettsiae posing an emerging global health risk, genetic requirements their infectious life cycles remain poorly understood. A major hurdle toward building this understanding has been lack efficient tools for manipulation, owing technical difficulties associated with obligate intracellular nature. To end, we implemented Tet-On system enable conditional gene expression in parkeri . Using Tet-On, show inducible antibiotic resistance fluorescent reporter. We further used promoter screen ability R. express four variants catalytically dead Cas9 (dCas9). demonstrate that all dCas9 can be expressed CRISPR interference (CRISPRi)-mediated targeted knockdown. knockdown as well endogenous virulence factor sca2 Altogether, have developed systems CRISPRi-mediated first time rickettsiae, laying groundwork more scalable, mechanistic investigations into cycles. IMPORTANCE The spotted fever group contains vector-borne pathogenic bacteria neglected threats public health. Due nature development manipulation stunted, molecular underpinnings lifecycle Here, expand toolkit by introducing These allow relatively easy rickettsial expression. effectiveness these disrupting cycle using CRISPRi deplete factor. will crucial comprehensive detailed biology pathogenesis.

Language: Английский

Rickettsia -Host-Tick Interactions: Knowledge Advances and Gaps DOI Creative Commons
Hwan Keun Kim

Infection and Immunity, Journal Year: 2022, Volume and Issue: 90(9)

Published: Aug. 22, 2022

Ticks are hematophagous ectoparasites capable of transmitting multiple human pathogens. Environmental changes have supported the expansion ticks into new geographical areas that become epicenters tick-borne diseases (TBDs).

Language: Английский

Citations

40
Rickettsia parkeri forms extensive, stable contacts with the rough endoplasmic reticulum DOI

Yamilex Acevedo‐Sánchez,

Patrick J. Woida, Caroline Anderson

et al.

The Journal of Cell Biology, Journal Year: 2025, Volume and Issue: 224(3)

Published: Jan. 7, 2025

Upon invasion into the host cell, a subset of bacterial pathogens resides exclusively in cytosol. While previous research revealed how they reshape plasma membrane during invasion, subvert immune response, and hijack cytoskeletal dynamics to promote their motility, it was unclear if these also interacted with organelles this crowded intracellular space. Here, we examined obligate pathogen Rickettsia parkeri interacts endoplasmic reticulum (ER), large dynamic organelle spread throughout cell. Using live-cell microscopy transmission focused-ion-beam scanning electron microscopy, show that R. forms extensive contacts rough ER are ∼55 nm apart cover more than half surface. Depletion ER-specific tethers VAPA VAPB reduced rickettsia–ER contacts, were localized around rickettsiae. Overall, our findings illuminate an interkingdom contact uniquely mediated by rickettsiae mimics some characteristics traditional sites.

Language: Английский

Citations

1
Pathogenic Rickettsia spp. as emerging models for bacterial biology DOI Creative Commons
Brandon Sit, Rebecca L. Lamason

Journal of Bacteriology, Journal Year: 2024, Volume and Issue: 206(2)

Published: Feb. 5, 2024

Our understanding of free-living bacterial models like

Language: Английский

Citations

8
Recent advances in genetic systems in obligate intracellular human-pathogenic bacteria DOI Creative Commons
Derek J. Fisher, Paul A. Beare

Frontiers in Cellular and Infection Microbiology, Journal Year: 2023, Volume and Issue: 13

Published: June 19, 2023

The ability to genetically manipulate a pathogen is fundamental discovering factors governing host-pathogen interactions at the molecular level and critical for devising treatment prevention strategies. While genetic "toolbox" many important bacterial pathogens extensive, approaches modifying obligate intracellular were classically limited due in part uniqueness of their obligatory lifestyles. Many researchers have confronted these challenges over past two half decades leading development multiple construct plasmid-bearing recombinant strains chromosomal gene inactivation deletion mutants, along with gene-silencing methods enabling study essential genes. This review will highlight seminal achievements recent developments (past 5 years) Anaplasma spp., Rickettsia Chlamydia Coxiella burnetii including progress being made still intractable Orientia tsutsugamushi. Alongside commentary strengths weaknesses various approaches, future research directions be discussed include C. that should utility other bacteria. Collectively, appears bright unraveling pathogenic mechanisms significant pathogens.

Language: Английский

Citations

13
Molecular detection and prevalence of Anaplasma and Rickettsia species in rodents captured from wildlife-human interfaces in Iringa and Morogoro regions, Tanzania DOI
Venance T. Msoffe,

Festo K. Mwombeki,

Claus Thomas

et al.

Mammalia, Journal Year: 2025, Volume and Issue: unknown

Published: May 7, 2025

Abstract Rickettsia and Anaplasma are Gram-negative, nonmotile, obligate intracellular bacteria in the order les. While various studies have been conducted on these threatening pathogens humans domestic animals, potential role of small mammals as reservoir zoonotic transmission is poorly understood Tanzania. This study investigated prevalence identity infecting captured at wildlife-human interfaces Iringa Morogoro, A total 371 blood samples were analyzed using conventional PCR DNA fragment sequencing, targeting approximately 424 bp 16S rRNA gene for 383 citrate synthase ( gltA ) . spp. was detected 9.7 %, 6.74 % samples. Rattus rattus exhibited highest both (21.43 %) (28.57 %). Two species A. bovis ovis three R. asembonensis , felis typhi confirmed through sequence analysis. highlights presence rodents, indicating rodents maintenance pathogens, especially interfaces, also reports first time

Language: Английский

Citations

0
Epidemiology and genetic characteristics of tick-borne bacteria in dromedary camels of the world DOI
Rachid Selmi,

Hanène Belkahia,

Alireza Sazmand

et al.

Acta Tropica, Journal Year: 2022, Volume and Issue: 234, P. 106599 - 106599

Published: July 8, 2022

Language: Английский

Citations

15
Microbial symbionts are shared between ants and their associated beetles DOI Creative Commons
Catalina Valdivia,

Justin A. Newton,

Christoph von Beeren

et al.

Environmental Microbiology, Journal Year: 2023, Volume and Issue: 25(12), P. 3466 - 3483

Published: Nov. 15, 2023

The transmission of microbial symbionts across animal species could strongly affect their biology and evolution, but our understanding patterns dynamics is limited. Army ants (Formicidae: Dorylinae) hundreds closely associated insect guest (myrmecophiles) can provide unique insights into interspecific symbiont sharing. Here, we compared the microbiota workers larvae army ant Eciton burchellii with those 13 myrmecophile beetle using 16S rRNA amplicon sequencing. We found that previously characterized specialized bacterial were largely absent from myrmecophiles, whose communities usually dominated by Rickettsia, Wolbachia, Rickettsiella and/or Weissella. Strikingly, different myrmecophiles often shared identical genotypes these common bacteria. Protein-coding gene sequences confirmed close relationship Weissella strains colonizing larvae, some several species. Unexpectedly, also similar to infecting dissimilar animals inhabiting very habitats: trout whales. Together, data show interacting share much microbiota, versatile inhabit possibly transmit a diverse range hosts environments.

Language: Английский

Citations

9
Cell-selective proteomics reveal novel effectors secreted by an obligate intracellular bacterial pathogen DOI Creative Commons
Allen G. Sanderlin, Hannah K. Margolis,

Abigail F. Meyer

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: July 18, 2024

Abstract Pathogenic bacteria secrete protein effectors to hijack host machinery and remodel their infectious niche. Rickettsia spp. are obligate intracellular that can cause life-threatening disease, but absolute dependence on the cell has impeded discovery of rickettsial targets. We implemented bioorthogonal non-canonical amino acid tagging (BONCAT) during R. parkeri infection selectively label, isolate, identify delivered into cell. As first use BONCAT in an bacterium, our screen more than doubles number experimentally validated for genus. The seven novel secreted factors (Srfs) we identified include -specific proteins unknown function localize cytoplasm, mitochondria, ER. further show one such effector, SrfD, interacts with Sec61 translocon. Altogether, work uncovers a diverse set previously uncharacterized lays foundation deeper exploration host-pathogen interface.

Language: Английский

Citations

3
The Ankyrin Repeat Protein RARP-1 Is a Periplasmic Factor That Supports Rickettsia parkeri Growth and Host Cell Invasion DOI
Allen G. Sanderlin, Ruth E. Hanna, Rebecca L. Lamason

et al.

Journal of Bacteriology, Journal Year: 2022, Volume and Issue: 204(7)

Published: June 21, 2022

Rickettsia spp. are obligate intracellular bacterial pathogens that have evolved a variety of strategies to exploit their host cell niche. However, the factors contribute this lifestyle poorly understood. Here, we show conserved ankyrin repeat protein RARP-1 supports parkeri infection. Specifically, promotes efficient entry and growth within cytoplasm, but it is not necessary for cell-to-cell spread or evasion autophagy. We further demonstrate secreted into cytoplasm by R. parkeri. Instead, resides in periplasm, identify several binding partners predicted work concert with during Altogether, our data reveal plays critical role rickettsial life cycle. IMPORTANCERickettsia pose growing threat human health. Nevertheless, strict reliance on niche has hindered investigation molecular mechanisms driving This study yields much-needed insight RARP-1, which across genus yet been functionally characterized. Earlier had suggested cytoplasm. results from periplasm important both invasion cells These as novel regulator

Language: Английский

Citations

12
Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection DOI Creative Commons
Hanna J. Laukaitis-Yousey,

Triston T. Cooper,

Chanakan Suwanbongkot

et al.

PLoS Pathogens, Journal Year: 2022, Volume and Issue: 18(12), P. e1011045 - e1011045

Published: Dec. 21, 2022

Since its recognition in 1994 as the causative agent of human flea-borne spotted fever, Rickettsia felis , has been detected worldwide over 40 different arthropod species. The cat flea, Ctenocephalides is a well-described biological vector R . Unique to insect-borne rickettsiae, can employ multiple routes infection including inoculation via salivary secretions and potentially infectious flea feces into skin vertebrate hosts. Yet, little known molecular interactions governing subsequent transmission While obligate intracellular nature rickettsiae hampered function large-scale mutagenesis strategies, studies have shown efficiency mariner -based transposon systems Rickettsiales. Thus, this study aimed assess genetic mutants model elucidate genes involved infection. A Himar1 transposase was used generate transformants, which genome sequencing revealed insertion near 3’ end sca1 Alterations expression resulted unique phenotypes. :: tn mutant portrayed enhanced growth kinetics compared wild-type during vitro culture, rickettsial loads were significantly reduced As consequence decreased within infected donor fleas, exhibited limited potential. use biologically relevant provides evidence defective phenotype associated with

Language: Английский

Citations

12