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

FEMS Microbiol Lett. 2016 Sep;363(17):fnw181. doi: 10.1093/femsle/fnw181. Epub 2016 Jul 26.

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.

Keywords: ADP-ribosyltransferase; NarE; Neisseria gonorrhoeae.

MeSH terms

  • ADP Ribose Transferases / chemistry
  • ADP Ribose Transferases / genetics*
  • ADP Ribose Transferases / metabolism*
  • Adenosine Diphosphate / metabolism*
  • Amino Acid Sequence
  • Biocatalysis
  • Blotting, Western
  • Codon, Initiator
  • Humans
  • Neisseria gonorrhoeae / enzymology
  • Neisseria gonorrhoeae / genetics
  • Neisseria gonorrhoeae / metabolism*
  • Promoter Regions, Genetic
  • Sequence Deletion*

Substances

  • Codon, Initiator
  • Adenosine Diphosphate
  • ADP Ribose Transferases
  • NarE protein, Neisseria meningitidis