The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion

Paula I. Rodas, A. Said Álamos-Musre, Francisca P. Álvarez1, Alejandro Escobar, Cecilia V. Tapia, Eduardo Osorio, Carolina Otero, Iván L. Calderón, Juan A. Fuentes, Fernando Gil, Daniel Paredes-Sabja, Myron Christodoulides

Resultado de la investigación: Research - revisión exhaustivaArticle

  • 1 Citas

Resumen

The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis. The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE-6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.

IdiomaEnglish
Número de artículofnw181
PublicaciónFEMS Microbiology Letters
Volumen363
Número de edición17
DOI
EstadoPublished - 1 sep 2016

Huella dactilar

Adenosine Diphosphate Ribose
Neisseria gonorrhoeae
Adenosine Diphosphate
Enzymes
Proteins
ADP Ribose Transferases
Agmatine
beta-Defensins
Initiator Codon
Genetic Promoter Regions
Western Blotting
Bacteria
Amino Acids
Polymerase Chain Reaction
Genes

Keywords

    ASJC Scopus subject areas

    • Microbiology
    • Molecular Biology
    • Genetics

    Citar esto

    Rodas, P. I., Álamos-Musre, A. S., Álvarez1, F. P., Escobar, A., Tapia, C. V., Osorio, E., ... Christodoulides, M. (2016). The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion. FEMS Microbiology Letters, 363(17), [fnw181]. DOI: 10.1093/femsle/fnw181
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    title = "The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion",
    abstract = "The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis. The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE-6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.",
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    author = "Rodas, {Paula I.} and Álamos-Musre, {A. Said} and Álvarez1, {Francisca P.} and Alejandro Escobar and Tapia, {Cecilia V.} and Eduardo Osorio and Carolina Otero and Calderón, {Iván L.} and Fuentes, {Juan A.} and Fernando Gil and Daniel Paredes-Sabja and Myron Christodoulides",
    year = "2016",
    month = "9",
    doi = "10.1093/femsle/fnw181",
    volume = "363",
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    The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion. / Rodas, Paula I.; Álamos-Musre, A. Said; Álvarez1, Francisca P.; Escobar, Alejandro; Tapia, Cecilia V.; Osorio, Eduardo; Otero, Carolina; Calderón, Iván L.; Fuentes, Juan A.; Gil, Fernando; Paredes-Sabja, Daniel; Christodoulides, Myron.

    En: FEMS Microbiology Letters, Vol. 363, N.º 17, fnw181, 01.09.2016.

    Resultado de la investigación: Research - revisión exhaustivaArticle

    TY - JOUR

    T1 - The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion

    AU - Rodas,Paula I.

    AU - Álamos-Musre,A. Said

    AU - Álvarez1,Francisca P.

    AU - Escobar,Alejandro

    AU - Tapia,Cecilia V.

    AU - Osorio,Eduardo

    AU - Otero,Carolina

    AU - Calderón,Iván L.

    AU - Fuentes,Juan A.

    AU - Gil,Fernando

    AU - Paredes-Sabja,Daniel

    AU - Christodoulides,Myron

    PY - 2016/9/1

    Y1 - 2016/9/1

    N2 - The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis. The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE-6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.

    AB - The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis. The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE-6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.

    KW - ADP-ribosyltransferase

    KW - NarE

    KW - Neisseria gonorrhoeae

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    U2 - 10.1093/femsle/fnw181

    DO - 10.1093/femsle/fnw181

    M3 - Article

    VL - 363

    JO - FEMS Microbiology Letters

    T2 - FEMS Microbiology Letters

    JF - FEMS Microbiology Letters

    SN - 0378-1097

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    Rodas PI, Álamos-Musre AS, Álvarez1 FP, Escobar A, Tapia CV, Osorio E y otros. The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion. FEMS Microbiology Letters. 2016 sep 1;363(17). fnw181. Disponible desde, DOI: 10.1093/femsle/fnw181