Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin
Bacteriocins synthesis is initiated from an inactive precursor, which is composed of an N-terminal leader peptide attached to a C-terminal pro-peptide. However, leaderless bacteriocins (LLB) do not possess this N-terminal leader peptide nor undergo post-translational modifications. These atypical ba...
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| Format: | Article |
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MDPI AG
2021-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/23/12877 |
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| author | Adrián Pérez-Ramos Rabia Ladjouzi Abdellah Benachour Djamel Drider |
| author_facet | Adrián Pérez-Ramos Rabia Ladjouzi Abdellah Benachour Djamel Drider |
| author_sort | Adrián Pérez-Ramos |
| collection | DOAJ |
| description | Bacteriocins synthesis is initiated from an inactive precursor, which is composed of an N-terminal leader peptide attached to a C-terminal pro-peptide. However, leaderless bacteriocins (LLB) do not possess this N-terminal leader peptide nor undergo post-translational modifications. These atypical bacteriocins are observed to be immediately active after their translation in the cytoplasm. However, although considered to be simple, the biosynthetic pathway of LLB remains to be fully understood. Enterocin DD14 (EntDD14) is a two-peptide LLB produced by <i>Enterococcus faecalis</i> 14, which is a strain isolated from meconium. In silico analysis of DNA encoding EntDD14 located a cluster of 10 genes <i>ddABCDEFGHIJ</i>, where <i>ddE</i> and <i>ddF</i> encode the peculiar DdE and DdF proteins, carrying pleckstrin homology (PH) domains. These modules are quite common in Eucarya proteins and are known to be involved in intracellular signaling or cytoskeleton organization. To elucidate their role within the EntDD14 genetic determinants, we constructed deletion mutants of the <i>ddE</i> and <i>ddF</i> genes. As a result, the mutants were unable to export EntDD14 outside of the cytoplasm even though there was a clear expression of structural genes <i>ddAB</i> encoding EntDD14, and genes <i>ddHIJ</i> encoding an ABC transporter. Importantly, in these mutant strains (Δ<i>ddE</i> and Δ<i>ddF</i>), EntDD14 was detected by mass spectrometry in the intracellular soluble fraction exerting, upon its accumulation, a toxic effect on the producing strain as revealed by cell-counting and confocal microscopy analysis. Taken together, these results clearly indicate that PH domain-containing proteins, such as DdE and DdF, are involved in the transport of the leaderless two-peptide EntDD14. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:53:28Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-971c79cba8824d098a0c909e0f264aa72023-11-23T02:29:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-11-0122231287710.3390/ijms222312877Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide BacteriocinAdrián Pérez-Ramos0Rabia Ladjouzi1Abdellah Benachour2Djamel Drider3UMR Transfrontalière BioEcoAgro 1158, University Lille, INRAE, University Liège, UPJV, YNCREA, University Artois, University Littoral Côte d’Opale, ICV—Institut Charles Viollette, 59000 Lille, FranceUMR Transfrontalière BioEcoAgro 1158, University Lille, INRAE, University Liège, UPJV, YNCREA, University Artois, University Littoral Côte d’Opale, ICV—Institut Charles Viollette, 59000 Lille, FranceUR Risques Microbiens, Normandie University, UNICAEN, U2RM, 14000 Caen, FranceUMR Transfrontalière BioEcoAgro 1158, University Lille, INRAE, University Liège, UPJV, YNCREA, University Artois, University Littoral Côte d’Opale, ICV—Institut Charles Viollette, 59000 Lille, FranceBacteriocins synthesis is initiated from an inactive precursor, which is composed of an N-terminal leader peptide attached to a C-terminal pro-peptide. However, leaderless bacteriocins (LLB) do not possess this N-terminal leader peptide nor undergo post-translational modifications. These atypical bacteriocins are observed to be immediately active after their translation in the cytoplasm. However, although considered to be simple, the biosynthetic pathway of LLB remains to be fully understood. Enterocin DD14 (EntDD14) is a two-peptide LLB produced by <i>Enterococcus faecalis</i> 14, which is a strain isolated from meconium. In silico analysis of DNA encoding EntDD14 located a cluster of 10 genes <i>ddABCDEFGHIJ</i>, where <i>ddE</i> and <i>ddF</i> encode the peculiar DdE and DdF proteins, carrying pleckstrin homology (PH) domains. These modules are quite common in Eucarya proteins and are known to be involved in intracellular signaling or cytoskeleton organization. To elucidate their role within the EntDD14 genetic determinants, we constructed deletion mutants of the <i>ddE</i> and <i>ddF</i> genes. As a result, the mutants were unable to export EntDD14 outside of the cytoplasm even though there was a clear expression of structural genes <i>ddAB</i> encoding EntDD14, and genes <i>ddHIJ</i> encoding an ABC transporter. Importantly, in these mutant strains (Δ<i>ddE</i> and Δ<i>ddF</i>), EntDD14 was detected by mass spectrometry in the intracellular soluble fraction exerting, upon its accumulation, a toxic effect on the producing strain as revealed by cell-counting and confocal microscopy analysis. Taken together, these results clearly indicate that PH domain-containing proteins, such as DdE and DdF, are involved in the transport of the leaderless two-peptide EntDD14.https://www.mdpi.com/1422-0067/22/23/12877leaderless bacteriocinsEntDD14pleckstrin homology domainbacteriocin transport |
| spellingShingle | Adrián Pérez-Ramos Rabia Ladjouzi Abdellah Benachour Djamel Drider Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin International Journal of Molecular Sciences leaderless bacteriocins EntDD14 pleckstrin homology domain bacteriocin transport |
| title | Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin |
| title_full | Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin |
| title_fullStr | Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin |
| title_full_unstemmed | Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin |
| title_short | Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin |
| title_sort | evidence for the involvement of pleckstrin homology domain containing proteins in the transport of enterocin dd14 entdd14 a leaderless two peptide bacteriocin |
| topic | leaderless bacteriocins EntDD14 pleckstrin homology domain bacteriocin transport |
| url | https://www.mdpi.com/1422-0067/22/23/12877 |
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