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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/217137
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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 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Congreso Journal http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
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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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eng |
language |
eng |
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info:eu-repo/semantics/altIdentifier/url/https://www.saib.org.ar/sites/default/files/TSP_BIOCELL_46213-SAIB-SAMIGE%202021.pdf |
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SAIB SAMIGE |
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SAIB SAMIGE |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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