Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer

Autores
Costa, M.; de Castro, Rosana Esther; Giménez, M. I.
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The halophilic archaeon Haloferax volcanii develops in a wide range of salinities (1.5-3.5 M NaCl) and, due to its ease of cultivation in the laboratory and the possibility of being genetically manipulated, it is used as a model organism for the study of archaeal biology. In the context of a comparative proteomics study of a rhomboid protease (RhoII) gene deletion mutant, we identified several proteins involved in metal homeostasis and cell surface structure/s assembly with differences in concentration and/or electrophoretic mobility in the protease deficient strain. Out of these, some were proteins which had not been previously characterized in H. volcanii, and that may constitute RhoII endogenous substrates. With the aim of understanding their physiological role, two of these annotated proteins (HVO_1153 and HVO_2150) were selected to generate and characterize the corresponding gene knock-out mutants. The hypothetical protein HVO_1153 primary sequence shows homology to adhesins and flagellins, and the hvo_2150 gene encodes HcpG, a predicted small copper protein (similar to plastocyanin and azurin) that may participate in electron transport and/or act as a copper reservoir in the cell membrane of H. volcanii. Genes were removed from the wild type chromosome by the ?pop-in / pop-out" method and the deletion in the null mutants was confirmed by PCR. The successful generation of the null mutant strains indicated that these genes are not essential for H. volcanii viability. The Δhvo_1153 mutant evidenced no differences in cell/colony morphology, cell adhesion to glass surfaces or growth in liquid medium at different conditions, when compared to the wild type strain. However, this mutant strain showed decreased motility in soft agar plates, in agreement with its predicted function in the databases. The ∆hcpGmutant did not exhibit deficiencies in growth in rich or minimal medium, when compared with the wild strain. Further phenotypic characterization of H. volcanii ∆hcpG is still ongoing. Altogether, the results presented in this work provide information regarding the role of two proteins which had not been previously characterized in H. volcanii and contribute to the general understanding of haloarchaeal physiology. Funded by ANPCyT and UNMdP.
Fil: Costa, M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Giménez, M. I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Congreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE)
Argentina
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
Sociedad Argentina de Microbiología General
Materia
HALOFERAX
MUTANTS
MOTILITY
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/217137

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network_name_str CONICET Digital (CONICET)
spelling Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron TransferCosta, M.de Castro, Rosana EstherGiménez, M. I.HALOFERAXMUTANTSMOTILITYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The halophilic archaeon Haloferax volcanii develops in a wide range of salinities (1.5-3.5 M NaCl) and, due to its ease of cultivation in the laboratory and the possibility of being genetically manipulated, it is used as a model organism for the study of archaeal biology. In the context of a comparative proteomics study of a rhomboid protease (RhoII) gene deletion mutant, we identified several proteins involved in metal homeostasis and cell surface structure/s assembly with differences in concentration and/or electrophoretic mobility in the protease deficient strain. Out of these, some were proteins which had not been previously characterized in H. volcanii, and that may constitute RhoII endogenous substrates. With the aim of understanding their physiological role, two of these annotated proteins (HVO_1153 and HVO_2150) were selected to generate and characterize the corresponding gene knock-out mutants. The hypothetical protein HVO_1153 primary sequence shows homology to adhesins and flagellins, and the hvo_2150 gene encodes HcpG, a predicted small copper protein (similar to plastocyanin and azurin) that may participate in electron transport and/or act as a copper reservoir in the cell membrane of H. volcanii. Genes were removed from the wild type chromosome by the ?pop-in / pop-out" method and the deletion in the null mutants was confirmed by PCR. The successful generation of the null mutant strains indicated that these genes are not essential for H. volcanii viability. The Δhvo_1153 mutant evidenced no differences in cell/colony morphology, cell adhesion to glass surfaces or growth in liquid medium at different conditions, when compared to the wild type strain. However, this mutant strain showed decreased motility in soft agar plates, in agreement with its predicted function in the databases. The ∆hcpGmutant did not exhibit deficiencies in growth in rich or minimal medium, when compared with the wild strain. Further phenotypic characterization of H. volcanii ∆hcpG is still ongoing. Altogether, the results presented in this work provide information regarding the role of two proteins which had not been previously characterized in H. volcanii and contribute to the general understanding of haloarchaeal physiology. Funded by ANPCyT and UNMdP.Fil: Costa, M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Giménez, M. I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaCongreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE)ArgentinaSociedad Argentina de Investigaciones en Bioquímica y Biología MolecularSociedad Argentina de Microbiología GeneralSAIB SAMIGE2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/217137Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer; Congreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE); Argentina; 2021; 113-1130327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saib.org.ar/sites/default/files/TSP_BIOCELL_46213-SAIB-SAMIGE%202021.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:15:24Zoai:ri.conicet.gov.ar:11336/217137instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:15:24.391CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
title Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
spellingShingle Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
Costa, M.
HALOFERAX
MUTANTS
MOTILITY
title_short Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
title_full Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
title_fullStr Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
title_full_unstemmed Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
title_sort Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer
dc.creator.none.fl_str_mv Costa, M.
de Castro, Rosana Esther
Giménez, M. I.
author Costa, M.
author_facet Costa, M.
de Castro, Rosana Esther
Giménez, M. I.
author_role author
author2 de Castro, Rosana Esther
Giménez, M. I.
author2_role author
author
dc.subject.none.fl_str_mv HALOFERAX
MUTANTS
MOTILITY
topic HALOFERAX
MUTANTS
MOTILITY
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The halophilic archaeon Haloferax volcanii develops in a wide range of salinities (1.5-3.5 M NaCl) and, due to its ease of cultivation in the laboratory and the possibility of being genetically manipulated, it is used as a model organism for the study of archaeal biology. In the context of a comparative proteomics study of a rhomboid protease (RhoII) gene deletion mutant, we identified several proteins involved in metal homeostasis and cell surface structure/s assembly with differences in concentration and/or electrophoretic mobility in the protease deficient strain. Out of these, some were proteins which had not been previously characterized in H. volcanii, and that may constitute RhoII endogenous substrates. With the aim of understanding their physiological role, two of these annotated proteins (HVO_1153 and HVO_2150) were selected to generate and characterize the corresponding gene knock-out mutants. The hypothetical protein HVO_1153 primary sequence shows homology to adhesins and flagellins, and the hvo_2150 gene encodes HcpG, a predicted small copper protein (similar to plastocyanin and azurin) that may participate in electron transport and/or act as a copper reservoir in the cell membrane of H. volcanii. Genes were removed from the wild type chromosome by the ?pop-in / pop-out" method and the deletion in the null mutants was confirmed by PCR. The successful generation of the null mutant strains indicated that these genes are not essential for H. volcanii viability. The Δhvo_1153 mutant evidenced no differences in cell/colony morphology, cell adhesion to glass surfaces or growth in liquid medium at different conditions, when compared to the wild type strain. However, this mutant strain showed decreased motility in soft agar plates, in agreement with its predicted function in the databases. The ∆hcpGmutant did not exhibit deficiencies in growth in rich or minimal medium, when compared with the wild strain. Further phenotypic characterization of H. volcanii ∆hcpG is still ongoing. Altogether, the results presented in this work provide information regarding the role of two proteins which had not been previously characterized in H. volcanii and contribute to the general understanding of haloarchaeal physiology. Funded by ANPCyT and UNMdP.
Fil: Costa, M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: de Castro, Rosana Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Giménez, M. I.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Congreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE)
Argentina
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
Sociedad Argentina de Microbiología General
description The halophilic archaeon Haloferax volcanii develops in a wide range of salinities (1.5-3.5 M NaCl) and, due to its ease of cultivation in the laboratory and the possibility of being genetically manipulated, it is used as a model organism for the study of archaeal biology. In the context of a comparative proteomics study of a rhomboid protease (RhoII) gene deletion mutant, we identified several proteins involved in metal homeostasis and cell surface structure/s assembly with differences in concentration and/or electrophoretic mobility in the protease deficient strain. Out of these, some were proteins which had not been previously characterized in H. volcanii, and that may constitute RhoII endogenous substrates. With the aim of understanding their physiological role, two of these annotated proteins (HVO_1153 and HVO_2150) were selected to generate and characterize the corresponding gene knock-out mutants. The hypothetical protein HVO_1153 primary sequence shows homology to adhesins and flagellins, and the hvo_2150 gene encodes HcpG, a predicted small copper protein (similar to plastocyanin and azurin) that may participate in electron transport and/or act as a copper reservoir in the cell membrane of H. volcanii. Genes were removed from the wild type chromosome by the ?pop-in / pop-out" method and the deletion in the null mutants was confirmed by PCR. The successful generation of the null mutant strains indicated that these genes are not essential for H. volcanii viability. The Δhvo_1153 mutant evidenced no differences in cell/colony morphology, cell adhesion to glass surfaces or growth in liquid medium at different conditions, when compared to the wild type strain. However, this mutant strain showed decreased motility in soft agar plates, in agreement with its predicted function in the databases. The ∆hcpGmutant did not exhibit deficiencies in growth in rich or minimal medium, when compared with the wild strain. Further phenotypic characterization of H. volcanii ∆hcpG is still ongoing. Altogether, the results presented in this work provide information regarding the role of two proteins which had not been previously characterized in H. volcanii and contribute to the general understanding of haloarchaeal physiology. Funded by ANPCyT and UNMdP.
publishDate 2022
dc.date.none.fl_str_mv 2022
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status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/217137
Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer; Congreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE); Argentina; 2021; 113-113
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/217137
identifier_str_mv Generation and Characterization of H. Volcanii Mutants en Genes with Predicted Roles in Motility and Electron Transfer; Congreso Conjunto SAIB-SAIGE 2021. LVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research (SAIB). XVI Annual Meeting of the Argentinean Society for General Microbiology (SAMIGE); Argentina; 2021; 113-113
0327-9545
1667-5746
CONICET Digital
CONICET
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