Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts

Amoeboflagellates of the genus Naegleria are free-living protists ubiquitously found in soil and freshwater habitats worldwide. They include the "brain-eating amoeba" Naegleria fowleri, an opportunistic pathogen that causes primary amoebic meningoencephalitis, a rare but fatal infection of...

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Autores: McGowan, Jamie, Kilias, Estelle S., Lipscombe, James, Alacid, Elisabet, Barker, Tom, Catchpole, Leah, McTaggart, Seanna, Warring, Sally D., Gharbi, Karim, Richards, Thomas A., Hall, Neil, Swarbreck, David
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/418654
Acesso em linha:http://hdl.handle.net/10261/418654
https://api.elsevier.com/content/abstract/scopus_id/105017668385
Access Level:acceso abierto
Palavra-chave:Heterolobosea
Legionellaceae
Amoeboflagellates
Effectors
Intracellular pathogens
Protists
Secretion systems
id ES_ffc402f577621cfe9154828fa48cb31d
oai_identifier_str oai:digital.csic.es:10261/418654
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
title Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
spellingShingle Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
McGowan, Jamie
Heterolobosea
Legionellaceae
Amoeboflagellates
Effectors
Intracellular pathogens
Protists
Secretion systems
title_short Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
title_full Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
title_fullStr Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
title_full_unstemmed Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
title_sort Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobionts
dc.creator.none.fl_str_mv McGowan, Jamie
Kilias, Estelle S.
Lipscombe, James
Alacid, Elisabet
Barker, Tom
Catchpole, Leah
McTaggart, Seanna
Warring, Sally D.
Gharbi, Karim
Richards, Thomas A.
Hall, Neil
Swarbreck, David
author McGowan, Jamie
author_facet McGowan, Jamie
Kilias, Estelle S.
Lipscombe, James
Alacid, Elisabet
Barker, Tom
Catchpole, Leah
McTaggart, Seanna
Warring, Sally D.
Gharbi, Karim
Richards, Thomas A.
Hall, Neil
Swarbreck, David
author_role author
author2 Kilias, Estelle S.
Lipscombe, James
Alacid, Elisabet
Barker, Tom
Catchpole, Leah
McTaggart, Seanna
Warring, Sally D.
Gharbi, Karim
Richards, Thomas A.
Hall, Neil
Swarbreck, David
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Wellcome Trust
Biotechnology and Biological Sciences Research Council (UK)
Royal Society (UK)
McGowan, Jamie [0000-0002-4240-5175]
dc.subject.none.fl_str_mv Heterolobosea
Legionellaceae
Amoeboflagellates
Effectors
Intracellular pathogens
Protists
Secretion systems
topic Heterolobosea
Legionellaceae
Amoeboflagellates
Effectors
Intracellular pathogens
Protists
Secretion systems
description Amoeboflagellates of the genus Naegleria are free-living protists ubiquitously found in soil and freshwater habitats worldwide. They include the "brain-eating amoeba" Naegleria fowleri, an opportunistic pathogen that causes primary amoebic meningoencephalitis, a rare but fatal infection of humans. Beyond their direct pathogenicity, protists can also act as environmental reservoirs for intracellular bacterial pathogens, such as Legionella spp., to persist and multiply in the environment. In this study, we carried out single-cell genome sequencing of two uncultivated Naegleria species isolated from the River Leam in England. From single cells, we generated two highly complete Naegleria genomes. Phylogenetic analyses placed these species as close relatives of Naegleria fultoni and Naegleria pagei. Exploring Naegleria evolutionary genomics, we identified gene families encoding antistasin-like domains, which have been characterized as factors that inhibit coagulation in blood-feeding leeches. Antistasin-like domains were identified in all sequenced Naegleria species and their close relative Willaertia magna, yet are otherwise largely restricted to animal genomes. Significantly, we recovered highly complete bacterial genomes from each Naegleria single-cell sample. Phylogenomic analysis revealed that both bacteria belong to the Legionellaceae family. Both bacterial genomes encode comprehensive sets of secretion systems and effector arsenals. We identified putative Legionella effectors that resemble TAL (Transcription activator-like) effectors from plant pathogenic Xanthomonas spp. in terms of protein sequence and predicted structure, representing a potentially novel class of Legionella effectors. Our study highlights the advantages of single-cell environmental genomics approaches, which enable direct association of intracellular pathogens with their hosts to better understand the evolution of host-pathogen interactions.IMPORTANCEBeyond their direct pathogenic potential, amoebae and other protists found in the environment can indirectly threaten human health by serving as reservoirs for intracellular bacterial pathogens to persist, evolve, and multiply in the environment. Despite their importance, protist-bacterial interactions remain poorly understood. In this study, we employed single-cell genomics to sequence the genomes of two uncultivated Naegleria amoebae, both harboring novel Legionella bacteria. From individual cells, we recovered highly complete eukaryotic and bacterial cobiont genome assemblies. Our work demonstrates the power of single-cell sequencing approaches in directly linking intracellular pathogens to their hosts to better understand the evolution of protist-bacterial interactions and the role that protists play in facilitating bacterial pathogens to persist long term in the environment.
publishDate 2025
dc.date.none.fl_str_mv 2025
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/418654
https://api.elsevier.com/content/abstract/scopus_id/105017668385
url http://hdl.handle.net/10261/418654
https://api.elsevier.com/content/abstract/scopus_id/105017668385
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Naegleria environmental single-cell sequencing. https://www.ebi.ac.uk/ena/browser/view/PRJEB85689
McGowan, Jamie; Kilias, Estelle S.; Lipscombe, James; Alacid, Elisabet; Barker, Tom; Catchpole, Leah; McTaggart, Seanna; Warring, Sally D.; Gharbi, Karim; Richards, Thomas A.; Hall, Neil; Swarbreck, David; 2025; Comparative single-cell genomics of two uncultivated Naegleria species harbouring Legionella cobionts [Dataset]; Zenodo; https://doi.org/10.5281/zenodo.15721720
https://doi.org/10.1128/msphere.00352-25
No
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Comparative single-cell genomics of two uncultivated Naegleria species harboring Legionella cobiontsMcGowan, JamieKilias, Estelle S.Lipscombe, JamesAlacid, ElisabetBarker, TomCatchpole, LeahMcTaggart, SeannaWarring, Sally D.Gharbi, KarimRichards, Thomas A.Hall, NeilSwarbreck, DavidHeteroloboseaLegionellaceaeAmoeboflagellatesEffectorsIntracellular pathogensProtistsSecretion systemsAmoeboflagellates of the genus Naegleria are free-living protists ubiquitously found in soil and freshwater habitats worldwide. They include the "brain-eating amoeba" Naegleria fowleri, an opportunistic pathogen that causes primary amoebic meningoencephalitis, a rare but fatal infection of humans. Beyond their direct pathogenicity, protists can also act as environmental reservoirs for intracellular bacterial pathogens, such as Legionella spp., to persist and multiply in the environment. In this study, we carried out single-cell genome sequencing of two uncultivated Naegleria species isolated from the River Leam in England. From single cells, we generated two highly complete Naegleria genomes. Phylogenetic analyses placed these species as close relatives of Naegleria fultoni and Naegleria pagei. Exploring Naegleria evolutionary genomics, we identified gene families encoding antistasin-like domains, which have been characterized as factors that inhibit coagulation in blood-feeding leeches. Antistasin-like domains were identified in all sequenced Naegleria species and their close relative Willaertia magna, yet are otherwise largely restricted to animal genomes. Significantly, we recovered highly complete bacterial genomes from each Naegleria single-cell sample. Phylogenomic analysis revealed that both bacteria belong to the Legionellaceae family. Both bacterial genomes encode comprehensive sets of secretion systems and effector arsenals. We identified putative Legionella effectors that resemble TAL (Transcription activator-like) effectors from plant pathogenic Xanthomonas spp. in terms of protein sequence and predicted structure, representing a potentially novel class of Legionella effectors. Our study highlights the advantages of single-cell environmental genomics approaches, which enable direct association of intracellular pathogens with their hosts to better understand the evolution of host-pathogen interactions.IMPORTANCEBeyond their direct pathogenic potential, amoebae and other protists found in the environment can indirectly threaten human health by serving as reservoirs for intracellular bacterial pathogens to persist, evolve, and multiply in the environment. Despite their importance, protist-bacterial interactions remain poorly understood. In this study, we employed single-cell genomics to sequence the genomes of two uncultivated Naegleria amoebae, both harboring novel Legionella bacteria. From individual cells, we recovered highly complete eukaryotic and bacterial cobiont genome assemblies. Our work demonstrates the power of single-cell sequencing approaches in directly linking intracellular pathogens to their hosts to better understand the evolution of protist-bacterial interactions and the role that protists play in facilitating bacterial pathogens to persist long term in the environment.We would like to acknowledge the members of the Technical Genomics Group at the Earlham Institute and note the specific contributions of Chris Watkins, Sacha Lucchini, Kendall Baker, and Neil Shearer. We also acknowledge the work delivered via the Research Computing Group at the Earlham Institute who manages and delivers High-Performance Computing. This work was funded by the Wellcome Trust Darwin Tree of Life Awards (218328 and 226458) and by the Biotechnology and Biological Sciences Research Council (BBSRC), part of UK Research and Innovation, through the Core Capability Grant (BB/CCG2220/1) at the Earlham Institute; the Earlham Institute Strategic Program Grant Decoding Biodiversity (BBX011089/1) and its constituent work packages (BBS/E/ER/230002A and BBS/E/ER/230002B). T.A.R. is supported by a Royal Society University Research Fellowship (URF/R/191005).Peer reviewedAmerican Society for MicrobiologyWellcome TrustBiotechnology and Biological Sciences Research Council (UK)Royal Society (UK)McGowan, Jamie [0000-0002-4240-5175]202620262025info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/418654https://api.elsevier.com/content/abstract/scopus_id/105017668385reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésNaegleria environmental single-cell sequencing. https://www.ebi.ac.uk/ena/browser/view/PRJEB85689McGowan, Jamie; Kilias, Estelle S.; Lipscombe, James; Alacid, Elisabet; Barker, Tom; Catchpole, Leah; McTaggart, Seanna; Warring, Sally D.; Gharbi, Karim; Richards, Thomas A.; Hall, Neil; Swarbreck, David; 2025; Comparative single-cell genomics of two uncultivated Naegleria species harbouring Legionella cobionts [Dataset]; Zenodo; https://doi.org/10.5281/zenodo.15721720https://doi.org/10.1128/msphere.00352-25Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4186542026-05-22T06:33:51Z
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