Acid stress response and protein induction in Campylobacter jejuni isolates with different acid tolerance.
BACKGROUND
During the transmission route from poultry to the human host, the major foodborne pathogen C. jejuni may experience many types of stresses, including low pH caused by different acids. However, not all strains are equally sensitive to the stresses. The aim of this study was to investigate the response to acid stress of three sequenced C. jejuni strains with different acid tolerances using HCl and acetic acid.


RESULTS
Two-dimensional gel electrophoresis was used for proteomic analysis and proteins were radioactively labelled with methionine to identify proteins only related to acid exposure. To allow added radioactive methionine to be incorporated into induced proteins, a modified chemically defined broth was developed with the minimal amount of methionine necessary for satisfactory growth of all strains. Protein spots were analyzed using image software and identification was done with MALDI-TOF-TOF. The most acid-sensitive isolate was C. jejuni 327, followed by NCTC 11168 and isolate 305 as the most tolerant. Overall, induction of five proteins was observed within the pI range investigated: 19 kDa periplasmic protein (p19), thioredoxin-disulfide (TrxB), a hypothetical protein Cj0706 (Cj0706), molybdenum cofactor biosynthesis protein (MogA), and bacterioferritin (Dps). Strain and acid type dependent differences in the level of response were observed. For strain NCTC 11168, the induced proteins and the regulator fur were analysed at the transcriptomic level using qRT-PCR. In this transcriptomic analysis, only up-regulation of trxB and p19 was observed.


CONCLUSIONS
A defined medium that supports the growth of a range of Campylobacter strains and suitable for proteomic analysis was developed. Mainly proteins normally involved in iron control and oxidative stress defence were induced during acid stress of C. jejuni. Both strain and acid type affected sensitivity and response.
Department of Food Science, Faculty of Sciences Copenhagen University, Rolighedsvej 30, 1958, Frederiksberg C, Denmark.
Tina
T
Birk
Monica Takamiya
MT
Wik
René
R
Lametsch
Susanne
S
Knøchel
D019342:Acetic Acid / Q000633:toxicity*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D016123:Campylobacter jejuni / Q000737:chemistry / Q000187:drug effects* / Q000254:growth & development / Q000502:physiology*; D003470:Culture Media / Q000737:chemistry; D004361:Drug Tolerance; D015180:Electrophoresis, Gel, Two-Dimensional; D006851:Hydrochloric Acid / Q000633:toxicity*; D007553:Isotope Labeling / Q000379:methods; D008715:Methionine / Q000378:metabolism; D020543:Proteome / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013312:Stress, Physiological
D001426:Bacterial Proteins; D003470:Culture Media; D020543:Proteome; D008715:Methionine; D019342:Acetic Acid; D006851:Hydrochloric Acid