Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei
In bacteria, the two-component system PhoPR plays an important role in regulating many genes related to phosphate uptake and metabolism. In Lacticaseibacillus paracasei inactivation of the response regulator PhoP results in increased resistance to arsenite [As(III)]. A comparative transcriptomic ana...
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Elsevier
2025-01-01
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| Series: | Current Research in Microbial Sciences |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666517425000197 |
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| author | Daniela Corrales Cristina Alcántara Dinoraz Vélez Vicenta Devesa Vicente Monedero Manuel Zúñiga |
| author_facet | Daniela Corrales Cristina Alcántara Dinoraz Vélez Vicenta Devesa Vicente Monedero Manuel Zúñiga |
| author_sort | Daniela Corrales |
| collection | DOAJ |
| description | In bacteria, the two-component system PhoPR plays an important role in regulating many genes related to phosphate uptake and metabolism. In Lacticaseibacillus paracasei inactivation of the response regulator PhoP results in increased resistance to arsenite [As(III)]. A comparative transcriptomic analysis revealed that the absence of PhoP has a strong effect on the transcriptome, with about 57.5 % of Lc. paracasei genes being differentially expressed, although only 92 of the upregulated genes and 23 of the downregulated genes reached a fold change greater than 2. Among them, the phnDCEB cluster, encoding a putative ABC phosphonate transporter and the acetate kinase encoding gene ackA (LCABL_01600) were downregulated tenfold and sevenfold, respectively. In vitro binding assays with selected PhoP-regulated genes showed that phosphorylation of PhoP stimulated its binding to the promoter regions of pstS (phosphate ABC transporter binding subunit), phnD and glnA glutamine synthetase) whereas no binding to the poxL (pyruvate oxidase) or ackA putative promoter regions was detected. This result identified for the first time three genes/operons belonging to the Pho regulon in a Lactobacillaceae species. Mapping of the reads obtained in the transcriptomic analysis revealed that transcription of ackA was severely diminished in the PhoP mutant after a hairpin structure located within the ackA coding region. Inactivation of phnD did not affect As(III) resistance whereas inactivation of ackA resulted in the same level of resistance as that observed in the PhoP mutant. These finding strongly suggests that PhoP mutant As(III) resistance is due to downregulation of ackA. Possible mechanisms of action are discussed. |
| format | Article |
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| institution | DOAJ |
| issn | 2666-5174 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Elsevier |
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| series | Current Research in Microbial Sciences |
| spelling | doaj-art-3ab3db2d9fdb465ea63d7c93865b3ee32025-08-20T03:10:42ZengElsevierCurrent Research in Microbial Sciences2666-51742025-01-01810035710.1016/j.crmicr.2025.100357Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracaseiDaniela Corrales0Cristina Alcántara1Dinoraz Vélez2Vicenta Devesa3Vicente Monedero4Manuel Zúñiga5Lactic Acid Bacteria and Probiotics Laboratory, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Agustín Escardino 7, 46980 Paterna, SpainLactic Acid Bacteria and Probiotics Laboratory, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Agustín Escardino 7, 46980 Paterna, SpainNext-generation Approaches for Integrative Food Toxicology group, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Catedràtic Agustín Escardino 7, 46980 Paterna, SpainNext-generation Approaches for Integrative Food Toxicology group, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Catedràtic Agustín Escardino 7, 46980 Paterna, SpainLactic Acid Bacteria and Probiotics Laboratory, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Agustín Escardino 7, 46980 Paterna, SpainLactic Acid Bacteria and Probiotics Laboratory, Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC), Av. Agustín Escardino 7, 46980 Paterna, Spain; Corresponding author.In bacteria, the two-component system PhoPR plays an important role in regulating many genes related to phosphate uptake and metabolism. In Lacticaseibacillus paracasei inactivation of the response regulator PhoP results in increased resistance to arsenite [As(III)]. A comparative transcriptomic analysis revealed that the absence of PhoP has a strong effect on the transcriptome, with about 57.5 % of Lc. paracasei genes being differentially expressed, although only 92 of the upregulated genes and 23 of the downregulated genes reached a fold change greater than 2. Among them, the phnDCEB cluster, encoding a putative ABC phosphonate transporter and the acetate kinase encoding gene ackA (LCABL_01600) were downregulated tenfold and sevenfold, respectively. In vitro binding assays with selected PhoP-regulated genes showed that phosphorylation of PhoP stimulated its binding to the promoter regions of pstS (phosphate ABC transporter binding subunit), phnD and glnA glutamine synthetase) whereas no binding to the poxL (pyruvate oxidase) or ackA putative promoter regions was detected. This result identified for the first time three genes/operons belonging to the Pho regulon in a Lactobacillaceae species. Mapping of the reads obtained in the transcriptomic analysis revealed that transcription of ackA was severely diminished in the PhoP mutant after a hairpin structure located within the ackA coding region. Inactivation of phnD did not affect As(III) resistance whereas inactivation of ackA resulted in the same level of resistance as that observed in the PhoP mutant. These finding strongly suggests that PhoP mutant As(III) resistance is due to downregulation of ackA. Possible mechanisms of action are discussed.http://www.sciencedirect.com/science/article/pii/S2666517425000197Lacticaseibacillus paracaseiArsenitePhosphateAcetate kinasePho regulon |
| spellingShingle | Daniela Corrales Cristina Alcántara Dinoraz Vélez Vicenta Devesa Vicente Monedero Manuel Zúñiga Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei Current Research in Microbial Sciences Lacticaseibacillus paracasei Arsenite Phosphate Acetate kinase Pho regulon |
| title | Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei |
| title_full | Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei |
| title_fullStr | Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei |
| title_full_unstemmed | Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei |
| title_short | Unveiling the role of the PhoP master regulator in arsenite resistance through ackA downregulation in Lacticaseibacillus paracasei |
| title_sort | unveiling the role of the phop master regulator in arsenite resistance through acka downregulation in lacticaseibacillus paracasei |
| topic | Lacticaseibacillus paracasei Arsenite Phosphate Acetate kinase Pho regulon |
| url | http://www.sciencedirect.com/science/article/pii/S2666517425000197 |
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