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.
authors.affiliation: Center for Integrative Medicine and Innovative Sciences, Facultad de Medicina, Universidad Andres Bello, Santiago, Chile paula.rodas@unab.cl.
authors.forename: Paula I
authors.initials: PI
authors.lastname: Rodas
authors.affiliation: Center for Integrative Medicine and Innovative Sciences, Facultad de Medicina, Universidad Andres Bello, Santiago, Chile.
authors.forename: A Said
authors.initials: AS
authors.lastname: Álamos-Musre
authors.affiliation: Center for Integrative Medicine and Innovative Sciences, Facultad de Medicina, Universidad Andres Bello, Santiago, Chile.
authors.forename: Francisca P
authors.initials: FP
authors.lastname: Álvarez
authors.affiliation: Instituto de Investigación en Ciencias Odontológicas, Facultad de Odontología, Universidad de Chile, Santiago, Chile.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Escobar
authors.affiliation: Laboratorio Clínica Dávila, Santiago, Chile Laboratorio de Micología Médica, Programa de Microbiología y Micología, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: Cecilia V
authors.initials: CV
authors.lastname: Tapia
authors.affiliation: Servicio de Ginecología y Obstetricia, Clínica Dávila, Santiago, Chile.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Osorio
authors.affiliation: Center for Integrative Medicine and Innovative Sciences, Facultad de Medicina, Universidad Andres Bello, Santiago, Chile.
authors.forename: Carolina
authors.initials: C
authors.lastname: Otero
authors.affiliation: Laboratorio de Genética y Patogénesis Bacteriana, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
authors.forename: Iván L
authors.initials: IL
authors.lastname: Calderón
authors.affiliation: Laboratorio de Genética y Patogénesis Bacteriana, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Fuentes
authors.affiliation: Laboratorio de Genética y Patogénesis Bacteriana, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
authors.forename: Fernando
authors.initials: F
authors.lastname: Gil
authors.affiliation: Microbiota-Host Interactions and Clostridia Research Group, Departamento de Ciencias Biológicas, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile Center for Bioinformatics and Integrative Biology, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
authors.forename: Daniel
authors.initials: D
authors.lastname: Paredes-Sabja
authors.affiliation: Neisseria Research Group, Molecular Microbiology, Sir Henry Wellcome Laboratories, Clinical and Experimental Sciences, University of Southampton Faculty of Medicine, Southampton, England.
authors.forename: Myron
authors.initials: M
authors.lastname: Christodoulides
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D055162:Biocatalysis; D015153:Blotting, Western; D018387:Codon, Initiator; D006801:Humans; D009344:Neisseria gonorrhoeae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
chemical_list: D018387:Codon, Initiator; D000244:Adenosine Diphosphate; D036002:ADP Ribose Transferases; C480854:NarE protein, Neisseria meningitidis
keywords: ADP-ribosyltransferase; NarE; Neisseria gonorrhoeae