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

The NarE protein of Neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminus deletion
The NarE protein of Neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminus deletion
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 to part of the coding sequence of 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 ?-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.
0378-1097
1-31
Rodas, Paula I.
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Alamos-Musre, A. Said
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Álvarez, Francisca
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Escobar, Alejandro
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Tapia, Cecilia V.
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Osorio, Eduardo
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Otero, Carolina
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Calderón, Iván L.
1f3a8bcf-80bd-43d4-859f-056b05412591
Fuentes, Juan A.
16f66ddf-9a40-4c70-a2f0-f60a2bb265cc
Gil, Fernando
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Paredes-Sabja, Daniel
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Christodoulides, Myron
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Rodas, Paula I.
1d6034a4-4c25-4b6f-90de-24c36afd2de5
Alamos-Musre, A. Said
1dbd0c82-98b6-44cf-b7ff-fbe6e075c1dd
Álvarez, Francisca
58cd8acb-8386-430b-8554-09ec3a74d32a
Escobar, Alejandro
21fa809a-b6d6-496d-8ead-bf15af1a7657
Tapia, Cecilia V.
ff745235-da62-445d-afa6-a874fc9d68b6
Osorio, Eduardo
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Otero, Carolina
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Calderón, Iván L.
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Fuentes, Juan A.
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Gil, Fernando
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Paredes-Sabja, Daniel
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Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078

Rodas, Paula I., Alamos-Musre, A. Said, Álvarez, Francisca, Escobar, Alejandro, Tapia, Cecilia V., Osorio, Eduardo, Otero, Carolina, Calderón, Iván L., Fuentes, Juan A., Gil, Fernando, Paredes-Sabja, Daniel and Christodoulides, Myron (2016) The NarE protein of Neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminus deletion. FEMS Microbiology Letters, 1-31. (doi:10.1093/femsle/fnw181). (PMID:27465490)

Record type: Article

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 to part of the coding sequence of 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 ?-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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Accepted/In Press date: 21 July 2016
e-pub ahead of print date: 26 July 2016
Organisations: Clinical & Experimental Sciences

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Local EPrints ID: 399381
URI: https://eprints.soton.ac.uk/id/eprint/399381
ISSN: 0378-1097
PURE UUID: e0dee692-b25c-4b0b-9527-c51e6927f16f

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Date deposited: 15 Aug 2016 12:54
Last modified: 29 Oct 2019 06:35

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Contributors

Author: Paula I. Rodas
Author: A. Said Alamos-Musre
Author: Francisca Álvarez
Author: Alejandro Escobar
Author: Cecilia V. Tapia
Author: Eduardo Osorio
Author: Carolina Otero
Author: Iván L. Calderón
Author: Juan A. Fuentes
Author: Fernando Gil
Author: Daniel Paredes-Sabja

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