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  1. Article ; Online: Editorial.

    Brom, Susana / Grohman, Elisabeth / Llosa, Matxalen

    Plasmid

    2020  Volume 113, Page(s) 102526

    Language English
    Publishing date 2020-08-01
    Publishing country United States
    Document type Editorial ; Introductory Journal Article
    ZDB-ID 282384-6
    ISSN 1095-9890 ; 0147-619X
    ISSN (online) 1095-9890
    ISSN 0147-619X
    DOI 10.1016/j.plasmid.2020.102526
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  2. Article ; Online: Recruitment of heterologous substrates by bacterial secretion systems for transkingdom translocation.

    Guzmán-Herrador, Dolores L / Fernández-Gómez, Andrea / Llosa, Matxalen

    Frontiers in cellular and infection microbiology

    2023  Volume 13, Page(s) 1146000

    Abstract: Bacterial secretion systems mediate the selective exchange of macromolecules between bacteria and their environment, playing a pivotal role in processes such as horizontal gene transfer or virulence. Among the different families of secretion systems, ... ...

    Abstract Bacterial secretion systems mediate the selective exchange of macromolecules between bacteria and their environment, playing a pivotal role in processes such as horizontal gene transfer or virulence. Among the different families of secretion systems, Type III, IV and VI (T3SS, T4SS and T6SS) share the ability to inject their substrates into human cells, opening up the possibility of using them as customized injectors. For this to happen, it is necessary to understand how substrates are recruited and to be able to engineer secretion signals, so that the transmembrane machineries can recognize and translocate the desired substrates in place of their own. Other factors, such as recruiting proteins, chaperones, and the degree of unfolding required to cross through the secretion channel, may also affect transport. Advances in the knowledge of the secretion mechanism have allowed heterologous substrate engineering to accomplish translocation by T3SS, and to a lesser extent, T4SS and T6SS into human cells. In the case of T4SS, transport of nucleoprotein complexes adds a bonus to its biotechnological potential. Here, we review the current knowledge on substrate recognition by these secretion systems, the many examples of heterologous substrate translocation by engineering of secretion signals, and the current and future biotechnological and biomedical applications derived from this approach.
    MeSH term(s) Humans ; Bacterial Secretion Systems/genetics ; Bacteria/metabolism ; Virulence ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Type IV Secretion Systems/metabolism
    Chemical Substances Bacterial Secretion Systems ; Bacterial Proteins ; Type IV Secretion Systems
    Language English
    Publishing date 2023-03-06
    Publishing country Switzerland
    Document type Journal Article ; Review ; Research Support, Non-U.S. Gov't
    ZDB-ID 2619676-1
    ISSN 2235-2988 ; 2235-2988
    ISSN (online) 2235-2988
    ISSN 2235-2988
    DOI 10.3389/fcimb.2023.1146000
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  3. Article ; Online: Bacterial injection machines: Evolutionary diverse but functionally convergent.

    Bleves, Sophie / Galán, Jorge E / Llosa, Matxalen

    Cellular microbiology

    2020  Volume 22, Issue 5, Page(s) e13157

    Abstract: Many human pathogens use Type III, Type IV, and Type VI secretion systems to deliver effectors into their target cells. The contribution of these secretion systems to microbial virulence was the main focus of a workshop organised by the International ... ...

    Abstract Many human pathogens use Type III, Type IV, and Type VI secretion systems to deliver effectors into their target cells. The contribution of these secretion systems to microbial virulence was the main focus of a workshop organised by the International University of Andalusia in Spain. The meeting addressed structure-function, substrate recruitment, and translocation processes, which differ widely on the different secretion machineries, as well as the nature of the translocated effectors and their roles in subverting the host cell. An excellent panel of worldwide speakers presented the state of the art of the field, highlighting the involvement of bacterial secretion in human disease and discussing mechanistic aspects of bacterial pathogenicity, which can provide the bases for the development of novel antivirulence strategies.
    Language English
    Publishing date 2020-01-16
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1468320-9
    ISSN 1462-5822 ; 1462-5814
    ISSN (online) 1462-5822
    ISSN 1462-5814
    DOI 10.1111/cmi.13157
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  4. Article: Coupling Proteins in Type IV Secretion.

    Llosa, Matxalen / Alkorta, Itziar

    Current topics in microbiology and immunology

    2018  Volume 413, Page(s) 143–168

    Abstract: Type IV coupling proteins (T4CPs) are essential constituents of most type IV secretion systems (T4SSs), and probably the most intriguing component in terms of their evolutionary origin and functional role. Coupling proteins have coevolved with their ... ...

    Abstract Type IV coupling proteins (T4CPs) are essential constituents of most type IV secretion systems (T4SSs), and probably the most intriguing component in terms of their evolutionary origin and functional role. Coupling proteins have coevolved with their cognate secretion system and translocated substrates. They are present in all conjugative systems, leading to the suggestion that they play a specific role in DNA transfer. However, they are also part of many T4SSs involved in bacterial virulence, where they are required for protein translocation, with no apparent involvement in DNA secretion. Their name reflects genetic and biochemical evidence of a connecting role between the substrate and the T4SS, thus probably playing a major role in substrate recruitment. Increasing evidence supports also a role in signal transmission leading to activation of secretion. Most studies have addressed conjugative coupling proteins of the VirD4-like protein family. Their conserved features include a nucleotide-binding domain, essential for substrate translocation, a C-terminal domain involved in substrate interactions, and a transmembrane domain anchoring them to the inner membrane, which is an important regulator of protein function. Purified soluble deletion mutants display ATP hydrolysis activity and unspecific DNA binding. Elucidation of the 3D structure of the soluble deletion mutant of the conjugative coupling protein TrwB, TrwBΔN70, provided the basis for further mutagenesis studies rendering interesting insights into the structure-function of these proteins. Their key role as couplers between substrate and transporter provides biotechnological potential as targets for anti-virulence strategies, as well as for customization of substrate delivery through heterologous secretion systems.
    MeSH term(s) Bacterial Proteins ; Conjugation, Genetic ; Protein Transport ; Type IV Secretion Systems/metabolism
    Chemical Substances Bacterial Proteins ; Type IV Secretion Systems
    Language English
    Publishing date 2018-03-13
    Publishing country Germany
    Document type Journal Article
    ISSN 0070-217X
    ISSN 0070-217X
    DOI 10.1007/978-3-319-75241-9_6
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  5. Article ; Online: The secret life of conjugative relaxases.

    Guzmán-Herrador, Dolores Lucía / Llosa, Matxalen

    Plasmid

    2019  Volume 104, Page(s) 102415

    Abstract: Conjugative relaxases are well-characterized proteins responsible for the site- and strand-specific endonucleolytic cleavage and strand transfer reactions taking place at the start and end of the conjugative DNA transfer process. Most of the relaxases ... ...

    Abstract Conjugative relaxases are well-characterized proteins responsible for the site- and strand-specific endonucleolytic cleavage and strand transfer reactions taking place at the start and end of the conjugative DNA transfer process. Most of the relaxases characterized biochemically and structurally belong to the HUH family of endonucleases. However, an increasing number of new families of relaxases are revealing a variety of protein folds and catalytic alternatives to accomplish conjugative DNA processing. Relaxases show high specificity for their cognate target DNA sequences, but several recent reports underscore the importance of their activity on secondary targets, leading to widespread mobilization of plasmids containing an oriT-like sequence. Some relaxases perform other functions associated with their nicking and strand transfer ability, such as catalyzing site-specific recombination or initiation of plasmid replication. They perform these roles in the absence of conjugation, and the validation of these functions in several systems strongly suggest that they are not mere artifactual laboratory observations. Other unexpected roles recently assigned to relaxases include controlling plasmid copy number and promoting retrotransposition. Their capacity to mediate promiscuous mobilization and genetic reorganizations can be exploited for a number of imaginative biotechnological applications. Overall, there is increasing evidence that conjugative relaxases are not only key enzymes for horizontal gene transfer, but may have been adapted to perform other roles which contribute to prokaryotic genetic plasticity. Relaxed target specificity may be key to this versatility.
    MeSH term(s) Bacteria/enzymology ; Bacteria/genetics ; Bacteria/metabolism ; Biotechnology ; Conjugation, Genetic ; DNA Replication ; DNA, Bacterial ; Endonucleases/chemistry ; Endonucleases/genetics ; Endonucleases/metabolism ; Plasmids/genetics ; Plasmids/metabolism ; Recombination, Genetic
    Chemical Substances DNA, Bacterial ; Endonucleases (EC 3.1.-)
    Language English
    Publishing date 2019-05-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 282384-6
    ISSN 1095-9890 ; 0147-619X
    ISSN (online) 1095-9890
    ISSN 0147-619X
    DOI 10.1016/j.plasmid.2019.102415
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  6. Article: Conjugative DNA Transfer From

    Samperio, Sara / Guzmán-Herrador, Dolores L / May-Cuz, Rigoberto / Martín, Maria Cruz / Álvarez, Miguel A / Llosa, Matxalen

    Frontiers in microbiology

    2021  Volume 12, Page(s) 606629

    Abstract: Lactic acid bacteria (LAB) belonging to the genus classically known ... ...

    Abstract Lactic acid bacteria (LAB) belonging to the genus classically known as
    Language English
    Publishing date 2021-02-11
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2021.606629
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  7. Article: The secret life of conjugative relaxases

    Guzmán-Herrador, Dolores Lucía / Llosa, Matxalen

    Plasmid. 2019 July, v. 104

    2019  

    Abstract: Conjugative relaxases are well-characterized proteins responsible for the site- and strand-specific endonucleolytic cleavage and strand transfer reactions taking place at the start and end of the conjugative DNA transfer process. Most of the relaxases ... ...

    Abstract Conjugative relaxases are well-characterized proteins responsible for the site- and strand-specific endonucleolytic cleavage and strand transfer reactions taking place at the start and end of the conjugative DNA transfer process. Most of the relaxases characterized biochemically and structurally belong to the HUH family of endonucleases. However, an increasing number of new families of relaxases are revealing a variety of protein folds and catalytic alternatives to accomplish conjugative DNA processing. Relaxases show high specificity for their cognate target DNA sequences, but several recent reports underscore the importance of their activity on secondary targets, leading to widespread mobilization of plasmids containing an oriT-like sequence. Some relaxases perform other functions associated with their nicking and strand transfer ability, such as catalyzing site-specific recombination or initiation of plasmid replication. They perform these roles in the absence of conjugation, and the validation of these functions in several systems strongly suggest that they are not mere artifactual laboratory observations. Other unexpected roles recently assigned to relaxases include controlling plasmid copy number and promoting retrotransposition. Their capacity to mediate promiscuous mobilization and genetic reorganizations can be exploited for a number of imaginative biotechnological applications. Overall, there is increasing evidence that conjugative relaxases are not only key enzymes for horizontal gene transfer, but may have been adapted to perform other roles which contribute to prokaryotic genetic plasticity. Relaxed target specificity may be key to this versatility.
    Keywords horizontal gene transfer ; plasmids ; plasticity ; site-specific recombination
    Language English
    Dates of publication 2019-07
    Publishing place Elsevier Inc.
    Document type Article
    Note NAL-AP-2-clean
    ZDB-ID 282384-6
    ISSN 1095-9890 ; 0147-619X
    ISSN (online) 1095-9890
    ISSN 0147-619X
    DOI 10.1016/j.plasmid.2019.102415
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  8. Article ; Online: Chloramphenicol Selection of IS10 Transposition in the cat Promoter Region of Widely Used Cloning Vectors.

    Coral González-Prieto / Leticia Agúndez / Matxalen Llosa

    PLoS ONE, Vol 10, Iss 9, p e

    2015  Volume 0138615

    Abstract: The widely used pSU8 family of cloning vectors is based on a p15A replicon and a chloramphenicol acetyltransferase (cat) gene conferring chloramphenicol resistance. We frequently observed an increase in the size of plasmids derived from these vectors. ... ...

    Abstract The widely used pSU8 family of cloning vectors is based on a p15A replicon and a chloramphenicol acetyltransferase (cat) gene conferring chloramphenicol resistance. We frequently observed an increase in the size of plasmids derived from these vectors. Analysis of the bigger molecular species shows that they have an IS10 copy inserted at a specific site between the promoter and the cat open reading frame. Promoter activity from both ends of IS10 has been reported, suggesting that the insertion events could lead to higher CAT production. Insertions were observed in certain constructions containing inserts that could lead to plasmid instability. To test the possibility that IS10 insertions were selected as a response to chloramphenicol selection, we have grown these constructs in the presence of different amounts of antibiotic and we observed that insertions arise promptly under higher chloramphenicol selective pressure. IS10 is present in many E. coli laboratory strains, so the possibility of insertion in constructions involving cat-containing vectors should be taken into account. Using lower chloramphenicol concentrations could solve this problem.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2015-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: Chloramphenicol Selection of IS10 Transposition in the cat Promoter Region of Widely Used Cloning Vectors.

    González-Prieto, Coral / Agúndez, Leticia / Llosa, Matxalen

    PloS one

    2015  Volume 10, Issue 9, Page(s) e0138615

    Abstract: The widely used pSU8 family of cloning vectors is based on a p15A replicon and a chloramphenicol acetyltransferase (cat) gene conferring chloramphenicol resistance. We frequently observed an increase in the size of plasmids derived from these vectors. ... ...

    Abstract The widely used pSU8 family of cloning vectors is based on a p15A replicon and a chloramphenicol acetyltransferase (cat) gene conferring chloramphenicol resistance. We frequently observed an increase in the size of plasmids derived from these vectors. Analysis of the bigger molecular species shows that they have an IS10 copy inserted at a specific site between the promoter and the cat open reading frame. Promoter activity from both ends of IS10 has been reported, suggesting that the insertion events could lead to higher CAT production. Insertions were observed in certain constructions containing inserts that could lead to plasmid instability. To test the possibility that IS10 insertions were selected as a response to chloramphenicol selection, we have grown these constructs in the presence of different amounts of antibiotic and we observed that insertions arise promptly under higher chloramphenicol selective pressure. IS10 is present in many E. coli laboratory strains, so the possibility of insertion in constructions involving cat-containing vectors should be taken into account. Using lower chloramphenicol concentrations could solve this problem.
    MeSH term(s) Anti-Bacterial Agents/pharmacology ; Chloramphenicol/pharmacology ; Chloramphenicol O-Acetyltransferase/genetics ; Chromosomes, Bacterial ; Cloning, Molecular ; DNA Transposable Elements/genetics ; Escherichia coli/drug effects ; Escherichia coli/genetics ; Genetic Vectors/genetics ; Plasmids/genetics ; Promoter Regions, Genetic/genetics ; Recombination, Genetic
    Chemical Substances Anti-Bacterial Agents ; DNA Transposable Elements ; Chloramphenicol (66974FR9Q1) ; Chloramphenicol O-Acetyltransferase (EC 2.3.1.28)
    Language English
    Publishing date 2015
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0138615
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  10. Article ; Online: The Conjugative Relaxase TrwC Promotes Integration of Foreign DNA in the Human Genome.

    González-Prieto, Coral / Gabriel, Richard / Dehio, Christoph / Schmidt, Manfred / Llosa, Matxalen

    Applied and environmental microbiology

    2017  Volume 83, Issue 12

    Abstract: Bacterial conjugation is a mechanism of horizontal DNA transfer. The relaxase TrwC of the conjugative plasmid R388 cleaves one strand of the transferred DNA at ... ...

    Abstract Bacterial conjugation is a mechanism of horizontal DNA transfer. The relaxase TrwC of the conjugative plasmid R388 cleaves one strand of the transferred DNA at the
    Language English
    Publishing date 2017-06-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 223011-2
    ISSN 1098-5336 ; 0099-2240
    ISSN (online) 1098-5336
    ISSN 0099-2240
    DOI 10.1128/AEM.00207-17
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