Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5

BACKGROUND: Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) tha...

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Autores: Denise, Hubert, Poot, Jacqueline, Jimenez, Maribel, Ambit, Audrey, Herrmann, Daland C, Vermeulen, Arno N, Coombs, Graham H, Mottram, Jeremy C
Tipo de recurso: artículo
Fecha de publicación:2006
País:España
Institución:Instituto de Salud Carlos III (ISCIII)
Repositorio:Repisalud
Idioma:inglés
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/7028
Acceso en línea:http://hdl.handle.net/20.500.12105/7028
Access Level:acceso abierto
Palabra clave:Amino Acid Sequence
Animals
Blotting, Southern
Cricetinae
Cysteine Endopeptidases
Dogs
Gene Deletion
Gene Expression Regulation, Enzymologic
Genome, Protozoan
Humans
Leishmania infantum
Leishmania mexicana
Leishmaniasis, Visceral
Mesocricetus
Molecular Sequence Data
Mutation
Protozoan Proteins
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
U937 Cells
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spelling Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5Denise, HubertPoot, JacquelineJimenez, MaribelAmbit, AudreyHerrmann, Daland CVermeulen, Arno NCoombs, Graham HMottram, Jeremy CAmino Acid SequenceAnimalsBlotting, SouthernCricetinaeCysteine EndopeptidasesDogsGene DeletionGene Expression Regulation, EnzymologicGenome, ProtozoanHumansLeishmania infantumLeishmania mexicanaLeishmaniasis, VisceralMesocricetusMolecular Sequence DataMutationProtozoan ProteinsReverse Transcriptase Polymerase Chain ReactionSequence Homology, Amino AcidU937 CellsBACKGROUND: Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) that was isolated from a naturally infected dog and then cloned. We found that JPCM5 has attributes that make it an excellent laboratory model; different stages of the parasite life cycle can be studied in vitro, it is accessible to genetic manipulation and it has retained its virulence. Furthermore, the L. infantum JPCM5 genome has now been fully sequenced. RESULTS: We have further focused our studies on LiCPA, the L. infantum homologue to L. mexicana cysteine peptidase CPA. LiCPA was found to share a high percentage of amino acid identity with CPA proteins of other Leishmania species. Two independent LiCPA-deficient promastigote clones (DeltaLicpa) were generated and their phenotype characterised. In contrast to L. mexicana CPA-deficient mutants, both clones of DeltaLicpa were found to have significantly reduced virulence in vitro and in vivo. Re-expression of just one LiCPA allele (giving DeltaLicpa::CPA) was sufficient to complement the reduced infectivity of both DeltaLicpa mutants for human macrophages, which confirms the importance of LiCPA for L. infantum virulence. In contrast, in vivo experiments did not show any virulence recovery of the re-expressor clone DeltaLicpaC1::CPA compared with the CPA-deficient mutant DeltaLicpaC1. CONCLUSION: The data suggest that CPA is not essential for replication of L. infantum promastigotes, but is important for the host-parasite interaction. Further studies will be necessary to elucidate the precise roles that LiCPA plays and why the re-expression of LiCPA in the DeltaLicpa mutants complemented the gene deletion phenotype only in in vitro and not in in vivo infection of hamsters.BioMed Central (BMC)20192019-01-3020062006-11-1320062006-11-13journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/20.500.12105/7028reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/70282026-06-12T12:43:37Z
dc.title.none.fl_str_mv Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
title Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
spellingShingle Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
Denise, Hubert
Amino Acid Sequence
Animals
Blotting, Southern
Cricetinae
Cysteine Endopeptidases
Dogs
Gene Deletion
Gene Expression Regulation, Enzymologic
Genome, Protozoan
Humans
Leishmania infantum
Leishmania mexicana
Leishmaniasis, Visceral
Mesocricetus
Molecular Sequence Data
Mutation
Protozoan Proteins
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
U937 Cells
title_short Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
title_full Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
title_fullStr Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
title_full_unstemmed Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
title_sort Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
dc.creator.none.fl_str_mv Denise, Hubert
Poot, Jacqueline
Jimenez, Maribel
Ambit, Audrey
Herrmann, Daland C
Vermeulen, Arno N
Coombs, Graham H
Mottram, Jeremy C
author Denise, Hubert
author_facet Denise, Hubert
Poot, Jacqueline
Jimenez, Maribel
Ambit, Audrey
Herrmann, Daland C
Vermeulen, Arno N
Coombs, Graham H
Mottram, Jeremy C
author_role author
author2 Poot, Jacqueline
Jimenez, Maribel
Ambit, Audrey
Herrmann, Daland C
Vermeulen, Arno N
Coombs, Graham H
Mottram, Jeremy C
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv
dc.subject.none.fl_str_mv Amino Acid Sequence
Animals
Blotting, Southern
Cricetinae
Cysteine Endopeptidases
Dogs
Gene Deletion
Gene Expression Regulation, Enzymologic
Genome, Protozoan
Humans
Leishmania infantum
Leishmania mexicana
Leishmaniasis, Visceral
Mesocricetus
Molecular Sequence Data
Mutation
Protozoan Proteins
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
U937 Cells
topic Amino Acid Sequence
Animals
Blotting, Southern
Cricetinae
Cysteine Endopeptidases
Dogs
Gene Deletion
Gene Expression Regulation, Enzymologic
Genome, Protozoan
Humans
Leishmania infantum
Leishmania mexicana
Leishmaniasis, Visceral
Mesocricetus
Molecular Sequence Data
Mutation
Protozoan Proteins
Reverse Transcriptase Polymerase Chain Reaction
Sequence Homology, Amino Acid
U937 Cells
description BACKGROUND: Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) that was isolated from a naturally infected dog and then cloned. We found that JPCM5 has attributes that make it an excellent laboratory model; different stages of the parasite life cycle can be studied in vitro, it is accessible to genetic manipulation and it has retained its virulence. Furthermore, the L. infantum JPCM5 genome has now been fully sequenced. RESULTS: We have further focused our studies on LiCPA, the L. infantum homologue to L. mexicana cysteine peptidase CPA. LiCPA was found to share a high percentage of amino acid identity with CPA proteins of other Leishmania species. Two independent LiCPA-deficient promastigote clones (DeltaLicpa) were generated and their phenotype characterised. In contrast to L. mexicana CPA-deficient mutants, both clones of DeltaLicpa were found to have significantly reduced virulence in vitro and in vivo. Re-expression of just one LiCPA allele (giving DeltaLicpa::CPA) was sufficient to complement the reduced infectivity of both DeltaLicpa mutants for human macrophages, which confirms the importance of LiCPA for L. infantum virulence. In contrast, in vivo experiments did not show any virulence recovery of the re-expressor clone DeltaLicpaC1::CPA compared with the CPA-deficient mutant DeltaLicpaC1. CONCLUSION: The data suggest that CPA is not essential for replication of L. infantum promastigotes, but is important for the host-parasite interaction. Further studies will be necessary to elucidate the precise roles that LiCPA plays and why the re-expression of LiCPA in the DeltaLicpa mutants complemented the gene deletion phenotype only in in vitro and not in in vivo infection of hamsters.
publishDate 2006
dc.date.none.fl_str_mv 2006
2006-11-13
2006
2006-11-13
2019
2019-01-30
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12105/7028
url http://hdl.handle.net/20.500.12105/7028
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv BioMed Central (BMC)
publisher.none.fl_str_mv BioMed Central (BMC)
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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