Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection

Since its recognition in 1994 as the causative agent of human flea-borne spotted fever, Rickettsia felis, has been detected worldwide in over 40 different arthropod species. The cat flea, Ctenocephalides felis, is a well-described biological vector of R. felis. Unique to insect-borne rickettsiae, R....

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Autores principales: Laukaitis, Hanna J., Cooper, Triston T., Suwanbongkot, Chanakan, Verhoeve, Victoria I., Kurtti, Timothy J., Munderloh, Ulrike G., Macaluso, Kevin R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9815595/
https://www.ncbi.nlm.nih.gov/pubmed/36542675
http://dx.doi.org/10.1371/journal.ppat.1011045
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author Laukaitis, Hanna J.
Cooper, Triston T.
Suwanbongkot, Chanakan
Verhoeve, Victoria I.
Kurtti, Timothy J.
Munderloh, Ulrike G.
Macaluso, Kevin R.
author_facet Laukaitis, Hanna J.
Cooper, Triston T.
Suwanbongkot, Chanakan
Verhoeve, Victoria I.
Kurtti, Timothy J.
Munderloh, Ulrike G.
Macaluso, Kevin R.
author_sort Laukaitis, Hanna J.
collection PubMed
description Since its recognition in 1994 as the causative agent of human flea-borne spotted fever, Rickettsia felis, has been detected worldwide in over 40 different arthropod species. The cat flea, Ctenocephalides felis, is a well-described biological vector of R. felis. Unique to insect-borne rickettsiae, R. felis can employ multiple routes of infection including inoculation via salivary secretions and potentially infectious flea feces into the skin of vertebrate hosts. Yet, little is known of the molecular interactions governing flea infection and subsequent transmission of R. felis. While the obligate intracellular nature of rickettsiae has hampered the function of large-scale mutagenesis strategies, studies have shown the efficiency of mariner-based transposon systems in Rickettsiales. Thus, this study aimed to assess R. felis genetic mutants in a flea transmission model to elucidate genes involved in vector infection. A Himar1 transposase was used to generate R. felis transformants, in which subsequent genome sequencing revealed a transposon insertion near the 3’ end of sca1. Alterations in sca1 expression resulted in unique infection phenotypes. While the R. felis sca1::tn mutant portrayed enhanced growth kinetics compared to R. felis wild-type during in vitro culture, rickettsial loads were significantly reduced during flea infection. As a consequence of decreased rickettsial loads within infected donor fleas, R. felis sca1::tn exhibited limited transmission potential. Thus, the use of a biologically relevant model provides evidence of a defective phenotype associated with R. felis sca1::tn during flea infection.
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spelling pubmed-98155952023-01-06 Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection Laukaitis, Hanna J. Cooper, Triston T. Suwanbongkot, Chanakan Verhoeve, Victoria I. Kurtti, Timothy J. Munderloh, Ulrike G. Macaluso, Kevin R. PLoS Pathog Research Article Since its recognition in 1994 as the causative agent of human flea-borne spotted fever, Rickettsia felis, has been detected worldwide in over 40 different arthropod species. The cat flea, Ctenocephalides felis, is a well-described biological vector of R. felis. Unique to insect-borne rickettsiae, R. felis can employ multiple routes of infection including inoculation via salivary secretions and potentially infectious flea feces into the skin of vertebrate hosts. Yet, little is known of the molecular interactions governing flea infection and subsequent transmission of R. felis. While the obligate intracellular nature of rickettsiae has hampered the function of large-scale mutagenesis strategies, studies have shown the efficiency of mariner-based transposon systems in Rickettsiales. Thus, this study aimed to assess R. felis genetic mutants in a flea transmission model to elucidate genes involved in vector infection. A Himar1 transposase was used to generate R. felis transformants, in which subsequent genome sequencing revealed a transposon insertion near the 3’ end of sca1. Alterations in sca1 expression resulted in unique infection phenotypes. While the R. felis sca1::tn mutant portrayed enhanced growth kinetics compared to R. felis wild-type during in vitro culture, rickettsial loads were significantly reduced during flea infection. As a consequence of decreased rickettsial loads within infected donor fleas, R. felis sca1::tn exhibited limited transmission potential. Thus, the use of a biologically relevant model provides evidence of a defective phenotype associated with R. felis sca1::tn during flea infection. Public Library of Science 2022-12-21 /pmc/articles/PMC9815595/ /pubmed/36542675 http://dx.doi.org/10.1371/journal.ppat.1011045 Text en © 2022 Laukaitis et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Laukaitis, Hanna J.
Cooper, Triston T.
Suwanbongkot, Chanakan
Verhoeve, Victoria I.
Kurtti, Timothy J.
Munderloh, Ulrike G.
Macaluso, Kevin R.
Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title_full Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title_fullStr Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title_full_unstemmed Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title_short Transposon mutagenesis of Rickettsia felis sca1 confers a distinct phenotype during flea infection
title_sort transposon mutagenesis of rickettsia felis sca1 confers a distinct phenotype during flea infection
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9815595/
https://www.ncbi.nlm.nih.gov/pubmed/36542675
http://dx.doi.org/10.1371/journal.ppat.1011045
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