Identification of Genes Associated with Sensitivity to Ultraviolet A (UVA) Irradiation by Transposon Mutagenesis of Vibrio parahaemolyticus

Miki Maetani-Yasui,Thi Kim Ngan Bui,Akira Takahashi,Yohsuke Kinouchi,Mutsumi Aihara,Kazuaki Mawatari,Masatake Akutagawa,Takahiro Emoto,Airi Honjo,Takaaki Shimohata,Takashi Uebanso

doi:10.3390/app10165549

Abstract

Ultraviolet (UV) irradiation is used to disinfect water and food and can be classified as UVA (detected at wavelengths 320–400 nm), UVB (280–320 nm), and UVC (<280 nm). We developed a method for UVA sterilization of equipment with a UVA-light-emitting diode (LED); however, a high rate of fluence was needed to promote pathogen inactivation. The aim of this study was to identify genes associated with UVA sensitivity with the goal of improving UVA-LED-mediated bactericidal activity. We constructed a transposon-mutant library of Vibrio parahaemolyticus and selected six mutants with high sensitivity to UVA irradiation. Genes associated with this phenotype include F-type H+-transporting ATPases (atp), as well as those involved in general secretion (gsp), and ubiquinone and terpenoid-quinone biosynthesis (ubi). Gene complementation resulted in decreased sensitivity to UVA-LED. The atp mutants had lower intracellular adenosine triphosphate (ATP) concentrations than the wild-type treatment, with 20 mM L-serine resulting in elevated ATP concentrations and decreased sensitivity to UVA-LED. The gsp mutants exhibited high levels of extracellular protein transport and the ubi mutants exhibited significantly different intracellular concentrations of ubiquinone-8. Taken together, our results suggest that the protein products of the atp, gsp, and ubi genes may regulate sensitivity to UVA irradiation.

Highlights

Vibrio parahaemolyticus is a Gram-negative marine bacterial species that can cause acute gastroenteritis manifested by diarrhea, headache, vomiting, nausea, and abdominal pain; this is typically observed in individuals who have consumed raw or undercooked seafood [1,2,3,4].V. parahaemolyticus-induced gastroenteritis is mediated by the type III secretion system and by enterotoxins, including thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) [5,6,7].Food poisoning associated with V. parahaemolyticus is a frequent occurrence worldwide [8,9].Appl
We recently reported that bacterial systems that were effective for repairing deoxyribonucleic acid (DNA) damage secondary to UVC irradiation were not effective against damage elicited by UVA-light-emitting diode (LED) irradiation [18]
Information provided by the National Center for Biotechnology Information, Kyoto Encyclopedia of Genes and Genomes, and RIMD facilitated the classification of these six mutant strains into three functional/orthologous groups

Summary

Introduction

Vibrio parahaemolyticus is a Gram-negative marine bacterial species that can cause acute gastroenteritis manifested by diarrhea, headache, vomiting, nausea, and abdominal pain; this is typically observed in individuals who have consumed raw or undercooked seafood [1,2,3,4].V. parahaemolyticus-induced gastroenteritis is mediated by the type III secretion system and by enterotoxins, including thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) [5,6,7].Food poisoning associated with V. parahaemolyticus is a frequent occurrence worldwide [8,9].Appl. Vibrio parahaemolyticus is a Gram-negative marine bacterial species that can cause acute gastroenteritis manifested by diarrhea, headache, vomiting, nausea, and abdominal pain; this is typically observed in individuals who have consumed raw or undercooked seafood [1,2,3,4]. V. parahaemolyticus-induced gastroenteritis is mediated by the type III secretion system and by enterotoxins, including thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) [5,6,7]. Food poisoning associated with V. parahaemolyticus is a frequent occurrence worldwide [8,9].

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