A genetic screen reveals a periplasmic copper chaperone required for nitrite reductase activity in pathogenic Neisseria

Freda E-C Jen; Karrera Y Djoko; Stephen J Bent; Christopher J Day; Alastair G McEwan; Michael P Jennings

doi:10.1096/fj.15-270751

A genetic screen reveals a periplasmic copper chaperone required for nitrite reductase activity in pathogenic Neisseria

FASEB J. 2015 Sep;29(9):3828-38. doi: 10.1096/fj.15-270751. Epub 2015 Jun 1.

Authors

Freda E-C Jen¹, Karrera Y Djoko¹, Stephen J Bent¹, Christopher J Day¹, Alastair G McEwan¹, Michael P Jennings²

Affiliations

¹ *Institute for Glycomics, Griffith University, Gold Coast Campus, Southport, Queensland, Australia; School of Chemistry Molecular Biosciences, University of Queensland, St. Lucia, Queensland, Australia; and Molecular and Biomedical Science, Faculty of Sciences, University of Adelaide, Adelaide, South Australia, Australia.
² *Institute for Glycomics, Griffith University, Gold Coast Campus, Southport, Queensland, Australia; School of Chemistry Molecular Biosciences, University of Queensland, St. Lucia, Queensland, Australia; and Molecular and Biomedical Science, Faculty of Sciences, University of Adelaide, Adelaide, South Australia, Australia m.jennings@griffith.edu.au.

PMID: 26031293
DOI: 10.1096/fj.15-270751

Abstract

Under conditions of low oxygen availability, Neisseria meningitidis and Neisseria gonorrhoeae are able to respire via a partial denitrification pathway in which nitrite is converted to nitrous oxide. In this process, nitrite reductase (AniA), a copper (Cu)-containing protein converts nitrite to NO, and this product is converted to nitrous oxide by nitric oxide reductase (NorB). NorB also confers protection against toxic NO, and so we devised a conditional lethal screen, using a norB mutant, to identify mutants that were resistant to nitrite-dependent killing. After random-deletion mutagenesis of N. meningitidis, this genetic screen identified a gene encoding a Cu chaperone that is essential for AniA function, AccA. Purified AccA binds one Cu (I) ion and also possesses a second binding site for Cu (II). This novel periplasmic Cu chaperone (AccA) appears to be essential for provision of Cu ions to AniA of pathogenic Neisseria to generate an active nitrite reductase. Apart from the Neisseria genus, AccA is distributed across a wide range of environmental Proteobacteria species.

Keywords: denitrification; gonorrhea; meningitis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antigens, Bacterial / genetics
Antigens, Bacterial / metabolism
Bacterial Outer Membrane Proteins / genetics
Bacterial Outer Membrane Proteins / metabolism
Copper / metabolism*
INDEL Mutation
Molecular Chaperones / genetics
Molecular Chaperones / metabolism*
Mutation*
Neisseria gonorrhoeae / genetics
Neisseria gonorrhoeae / metabolism*
Neisseria meningitidis / genetics
Neisseria meningitidis / metabolism*
Nitrite Reductases / genetics
Nitrite Reductases / metabolism*
Periplasmic Proteins / genetics
Periplasmic Proteins / metabolism*

Substances

Antigens, Bacterial
Bacterial Outer Membrane Proteins
Molecular Chaperones
Periplasmic Proteins
aniA protein, Neisseria gonorrhoeae
Copper
Nitrite Reductases