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: Article

2 Citas (Scopus)

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.

Idioma originalEnglish
Número de artículofnw181
PublicaciónFEMS Microbiology Letters
Volumen363
N.º17
DOI
EstadoPublished - 1 sep 2016

Huella dactilar

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

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]. https://doi.org/10.1093/femsle/fnw181
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. / The NarE protein of neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion. En: FEMS Microbiology Letters. 2016 ; Vol. 363, N.º 17.
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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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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: Article

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

AN - SCOPUS:84987665873

VL - 363

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 17

M1 - fnw181

ER -