Alkali stress is an important means of inactivating undesirable pathogens in a wide range of situations. Unfortunately, Listeria monocytogenes can launch an alkaline tolerance response, significantly increasing persistence of the pathogen in such environments. This study compared transcriptome patterns of alkali and nonalkali-stressed L. monocytogenes 10403S cells, to elucidate the mechanisms by which Listeria adapts and/or grows during short- or long-term alkali stress. Transcription profiles associated with alkali shock (AS) were obtained by DNA microarray analysis of midexponential cells suspended in pH 9 media for 15, 30, or 60 min. Transcription profiles associated with alkali adaptation (AA) were obtained similarly from cells grown to midexponential phase at pH 9. Comparison of AS and AA transcription profiles with control cell profiles identified a high number of differentially regulated open-reading frames in all tested conditions. Rapid (15 min) changes in expression included upregulation of genes encoding for multiple metabolic pathways (including those associated with Na+/H+ antiporters), ATP-binding cassette transporters of functional compatible solutes, motility, and virulence-associated genes as well as the σB controlled stress resistance network. Slower (30 min and more) responses to AS and adaptation during growth in alkaline conditions (AA) involved a different pattern of changes in mRNA concentrations, and genes involved in proton export.
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References
1.
AbramsonJ, KabackHR, IwataS. Structural comparison of lactose permease and the glycerol-3-phosphate antiporter: members of the major facilitator superfamily. Curr Opin Struct Biol, 2004; 14:413–419.
2.
BealesN. Adaptation of microorganisms to cold temperatures, weak acid preservatives, low pH, and osmotic stress: a review. Compr Rev Food Sci Food Saf, 2004; 3:1–20.
3.
BierneH, CossartP. Listeria monocytogenes surface proteins: from genome predictions to function. Microbiol Mol Biol R, 2007; 71:377–397.
4.
BlankenhornD, PhillipsJ, SlonczewskiJL. Acid and base induced proteins during aerobic and anaerobic growth of Escherichia coli revealed by two-dimensional gel electrophoresis. J Bacteriol, 1999; 181:2209–2216.
5.
BoothIR, PourkomailianB, McLagganD, KooSP. Mechanisms controlling compatible solute accumulation—a consideration of the genetics and physiology of bacterial osmoregulation. J Food Eng, 1994; 22:381–397.
6.
ButlerT, FrenckRW, JohnsonRB, KhakhriaR. In vitro effects of azithromycin on Salmonella typhi: early inhibition by concentrations less than the MIC and reduction of MIC by alkaline pH and small inocula. J Antimicrob Chemother, 2001; 47:455–458.
7.
ChengJB, GuffantiAA, KrulwichTA. A two-gene ABC type transport system that extrudes Na+ in Bacillus subtilis is induced by ethanol or protonophore. Mol Microbiol, 1997; 23:1107–1120.
8.
Cheroutre-VialetteM, LebertI, HebraudM, LabadieJC, LebertA. Effects of pH or aw stress on growth of Listeria monocytogenes. Int J Food Microbiol, 1998; 42:71–77.
9.
ConteMP, PetroneG, Di BiaseAM, LonghiC, PentaM, TinariA, SupertiF, FabozziG, ViscaP, SegantiL. Effect of acid adaptation on the fate of Listeria monocytogenes in THP-1 human macrophages activated by gamma interferon. Infect Immun, 2002; 70:4369–4378.
10.
CotterPD, GahanCGM, HillC. Analysis of the role of the Listeria monocytogenes F0F1-ATPase operon in the acid tolerance response. Int J Food Microbiol, 2000; 60:137–146.
11.
CotterPD, RyanS, GahanCGM, HillC. Presence of GadD1 glutamate decarboxylase in selected Listeria monocytogenes strains is associated with an ability to grow at low pH. Appl Environ Microbiol, 2005; 71:2832–2839.
12.
DeutscherJ, GalinierA, Martin-VerstraeteI. Carbohydrate uptake and metabolism. Bacillus Subtilis and Its Closest Relatives: From Genes to Cells. SonensheinAL, HochJA, LosickR. Washington, DC: ASM Press, 2002; 129–150.
13.
DobsonCM. Folding and Binding. Curr Opin Struct Biol, 1993; 3:57–59.
14.
EdgarR, DomrachevM, LashAE. Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res, 2002; 30:207–210.
15.
EdmanDC, PollockMB, HallER. Listeria monocytogenes L forms. I. Induction maintenance, and biological characteristics. J Bacteriol, 1968; 96:352–357.
FerreiraA, GrayM, WiedmannM, BoorKJ. Comparative genomic analysis of the sigB operon in Listeria monocytogenes and in other gram-positive bacteria. Curr Microbiol, 2004; 48:39–46.
18.
FolioP, ChavantP, ChafseyI, BelkorchiaA, ChambonC, HebraudM. Two-dimensional electrophoresis database of Listeria monocytogenes EGDe proteome and proteomic analysis of mid-log and stationary growth phase cells. Proteomics, 2004; 4:3187–3201.
19.
FraserKR, HarvieD, CootePJ, O'ByrneCP. Identification and characterization of an ATP binding cassette L-carnitine transporter in Listeria monocytogenes. Appl Environ Microbiol, 2000; 66:4696–4704.
20.
FraserKR, SueD, WiedmannM, BoorK, O'ByrneCP. Role of sigma(B) in regulating the compatible solute uptake systems of Listeria monocytogenes: osmotic induction of opuC is sigma(B) dependent. Appl Environ Microbiol, 2003; 69:2015–2022.
21.
FujisawaM, WadaY, ItoM. Modulation of the K+ efflux activity of Bacillus subtilis YhaU by YhaT and the C-terminal region of YhaS. FEMS Microbiol Lett, 2004; 231:211–217.
22.
GahanCGM, O'MahonyJ, HillC. Characterization of the groESL operon in Listeria monocytogenes: utilization of two reporter systems (gfp and hly) for evaluating in vivo expression. Infect Immun, 2001; 69:3924–3932.
23.
GaoH, WangY, LiuX, YanT, WuL, AlmE, ArkinA, ThompsonDK, ZhouJ. Global transcriptome analysis of the heat shock response of Shewanella oneidensis. J Bacteriol, 2004; 186:7796–7803.
24.
GarnerMR, NjaaBL, WiedmannM, BoorKJ. Sigma B contributes to Listeria monocytogenes gastrointestinal infection, but not to systemic spread, in the guinea pig infection model. Infect Immun, 2006; 74:876–886.
25.
GeourjonC, OrelleC, SteinfelsE, BlanchetC, DeleageG, Di PietroA, JaultJM. A common mechanism for ATP hydrolysis in ABC transporter and helicase superfamilies. Trends Biochem Sci, 2001; 26:539–544.
26.
GerhardtPN, SmithLT, SmithGM. Sodium-driven, osmotically activated glycine betaine transport in Listeria monocytogenes membrane vesicles. J Bacteriol, 1996; 178:6105–6109.
27.
GerhardtPNM, SmithLT, SmithGM. Osmotic and chill activation of glycine betaine porter II in Listeria monocytogenes membrane vesicles. J Bacteriol, 2000; 182:2544–2550.
28.
GiotisES, BlairIS, McDowellDA. Effects of short term alkaline adaptation on surface properties of Listeria monocytogenes 10403S. Open Food Sci J, 2009; 4:62–65.
29.
GiotisES, BlairIS, McDowellDA. Morphological changes in Listeria monocytogenes subjected to sublethal alkaline stress. Int J Food Microbiol, 2007a. 120:250–258.
30.
GiotisES, McDowellDA, BlairIS, WilkinsonBJ. Role of branched-chain fatty acids in pH stress tolerance in Listeria monocytogenes. Appl Environ Microbiol, 2007b. 73:997–1001.
31.
GiotisES, MudchareeJ, WilkinsonBJ, BlairIS, McDowellDA. Role of sigB factor in the alkaline tolerance response (AlTR) of Listeria monocytogenes 10403S and cross protection to subsequent ethanol and osmotic stress. J Food Prot, 2008a. 71:1481–1485.
32.
GiotisES, MuthaiyanA, BlairIS, WilkinsonBJ, McDowellDA. Genomic and proteomic analysis of the alkali-tolerance response (AlTR) in Listeria monocytogenes 10403S. BMC Microbiol, 2008b. 8:38.
33.
HanawaT, FukudaM, KawakamiH, HiranoH, KamiyaS, YamamotoT. The Listeria monocytogenes DnaK chaperone is required for stress tolerance and efficient phagocytosis with macrophages. Cell Stress Chaperones, 1999; 4:118–128.
34.
HaseCC, FedorovaND, GalperinMY, DibrovPA. Sodium ion cycle in bacterial pathogens: evidence from cross genome comparisons. Microbiol Mol Biol Rev, 2001; 65:353–370.
35.
ItoM, XuHX, GuffantiAA, WeiY, ZviL, ClaphamDE, KrulwichTA. The voltage-gated Na+ channel NavBP has a role in motility, chemotaxis, and pH homeostasis of an alkaliphilic Bacillus. Proc Natl Acad Sci USA, 2004; 101:10566–10571.
36.
KarlinS, TheriotJ, MrazekJ. Comparative analysis of gene expression among low G-C gram-positive genomes. Proc Natl Acad Sci USA, 2004; 101:6182–6187.
37.
KathariouS. Listeria monocytogenes virulence and pathogenicity, a food safety perspective. J Food Prot, 2002; 65:1811–1829.
38.
KimH, MarquisH, BoorKJ. sigma(B) contributes to Listeria monocytogenes invasion by controlling expression of inlA and inlB. Microbiology (UK), 2005; 151:3215–3222.
39.
KrulwichTA. Alkaliphiles—basic molecular problems of pH tolerance and bioenergetics. Mol Microbiol, 1995; 15:403–410.
40.
LinTL, ConnellyMB, AmoloC, OtaniS, YaverDS. Global transcriptional response of Bacillus subtilis to treatment with subinhibitory concentrations of antibiotics that inhibit protein synthesis. Antimicrob Agents Chemother, 2005; 49:1915–1926.
41.
LiuSQ, GrahamJE, BigelowL, MorsePD, WilkinsonBJ. Identification of Listeria monocytogenes genes expressed in response to growth at low temperature. Appl Environ Microbiol, 2002; 68:1697–1705.
MaurerLM, YohannesE, BondurantSS, RadmacherM, SlonczewskiJL. pH regulates genes for flagellar motility, catabolism, and oxidative stress in Escherichia coli K-12. J Bacteriol, 2005; 187:304–319.
44.
MendumML, SmithLT. Gbu glycine betaine porter and carnitine uptake in osmotically stressed Listeria monocytogenes cells. Appl Environ Microbiol, 2002; 68:5647–5655.
45.
MoorheadSM, DykesGA. Influence of the sigB gene on the cold stress survival and subsequent recovery of two Listeria monocytogenes serotypes. Int J Food Microbiol, 2004; 91:63–72.
46.
NelsonKE, FoutsDE, MongodinEF, RavelJ, DeBoyRT, KolonayJF, RaskoDA, AngiuoliSV, GillSR, PaulsenIT, PetersonJ, WhiteO, NelsonWC, NiermanW, BeananMJ, BrinkacLM, DaughertySC, DodsonRJ, DurkinAS, MadupuR, HaftDH, SelengutJ, Van AkenS, KhouriH, FedorovaN, ForbergerH, TranB, KathariouS, WonderlingLD, UhlichGA, BaylesDO, LuchanskyJB, FraserCM. Whole genome comparisons of serotype 4b and 1/2a strains of the food-borne pathogen Listeria monocytogenes reveal new insights into the core genome components of this species. Nucleic Acids Res, 2004; 32:2386–2395.
47.
O'DriscollB. Characterisation of the Acid Tolerance Response in Listeria monocytogenes. Galway, Ireland: National University of Ireland, 1997.
48.
OlierM, RousseauxS, PiveteauP, LemaitreJP, RoussetA, GuzzoJ. Screening of glutamate decarboxylase activity and bile salt resistance of human asymptomatic carriage, clinical, food, and environmental isolates of Listeria monocytogenes. Int J Food Microbiol, 2004; 93:87–99.
49.
PadanE, BibiE, ItoM, KrulwichTA. Alkaline pH homeostasis in bacteria: new insights. Biochim Biophys Acta Biomembr, 2005; 1717:67–88.
50.
RauchM, LuoQ, Muller-AltrockS, GoebelW. SigB dependent in vitro transcription of prfA and some newly identified genes of Listeria monocytogenes whose expression is affected by PrfA in vivo. J Bacteriol, 2005; 187:800–804.
51.
RiordanJT, MuthaiyanA, VoorhiesV, PriceW, ChristopherT, GrahamJE, WilkinsonBJ, GustafsonJE. Response of Staphylococcus aureus to salicylate challenge. J Bacteriol, 2007; 189:220–227.
52.
SatorhelyiP. Microarray Analyse der pH Stressantwort von Listeria monocytogenes und Corynebacterium glutamicum. München: Technischen Universität Eingereicht und Durch, 2004.
53.
SegalAW, GeisowM, GarciaR, HarperA, MillerR. The respiratory burst of phagocytic cells is associated with a rise in vacuolar pH. Nature, 1981; 290:406–409.
54.
SharmaM, TaorminaPJ, BeuchatLR. Habituation of foodborne pathogens exposed to extreme pH conditions: genetic basis and implications in foods and food processing environments. Food Sci Technol Res, 2003; 9:115–127.
55.
SleatorRD, HillC. A novel role for the LisRK two-component regulatory system in listerial osmotolerance. Clin Microbiol Infect, 2005; 11:599–601.
56.
SleatorRD, Wemekamp-KamphuisHH, GahanCGM, AbeeT, HillC. A PrfA-regulated bile exclusion system (BilE) is a novel virulence factor in Listeria monocytogenes. Mol Microbiol, 2005; 55:1183–1195.
57.
SueD, FinkD, WiedmannM, BoorKJ. Sigma(B)-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment. Microbiology (UK), 2004; 150:3843–3855.
58.
TakamiH, TakakiY, UchiyamaI. Genome sequence of Oceanobacillus iheyensis isolated from the Iheya Ridge and its unexpected adaptive capabilities to extreme environments. Nucleic Acids Res, 2002; 30:3927–3935.
59.
TaorminaPJ, BeuchatLR. Survival and growth of alkali-stressed Listeria monocytogenes on beef frankfurters and thermotolerance in frankfurter exudates. J Food Prot, 2002a. 65:291–298.
60.
TaorminaPJ, BeuchatLR. Survival of Listeria monocytogenes in commercial food-processing equipment cleaning solutions and subsequent sensitivity to sanitizers and heat. J Appl Microbiol, 2002b. 92:71–80.
61.
TusherVG, TibshiraniR, ChuG. Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci, 2001; 98:5116–5121.
62.
VasseurC, RigaudN, HebraudM, LabadieJ. Combined effects of NaCl, NaOH, and biocides (monolaurin or lauric acid) on inactivation of Listeria monocytogenes and Pseudomonas spp. J Food Prot, 2001; 64:1442–1445.
63.
WelshDT. Ecological significance of compatible solute accumulation by micro-organisms: from single cells to global climate. FEMS Microbiol Rev, 2000; 24:263–290.
64.
Wemekamp-KamphuisHH, WoutersJA, de LeeuwP, HainT, ChakrabortyT, AbeeT. Identification of sigma factor sigma(B)-controlled genes and their impact on acid stress, high hydrostatic pressure, and freeze survival in Listeria monocytogenes EGD-e. Appl Environ Microbiol, 2004; 70:3457–3466.
65.
WiedmannM, BruceJL, KeatingC, JohnsonAE, McDonoughPL, BattCA. Ribotypes and virulence gene polymorphisms suggest three distinct Listeria monocytogenes lineages with differences in pathogenic potential. Infect Immun, 1997; 65:2707–2716.
66.
WilhelmJ, HahnM, PingoudA. Influence of DNA target melting behavior on real-time PCR quantification. Clin Chem, 2000; 46:1738–1743.
WonderlingLD, WilkinsonBJ, BaylesDO. The htrA (degP) gene of Listeria monocytogenes 10403S is essential for optimal growth under stress conditions. Appl Environ Microbiol, 2004; 70:1935–1943.
69.
WoodJM, BremerE, CsonkaLN, KraemerR, PoolmanB, van der HeideT, SmithLT. Osmosensing and osmoregulatory compatible solute accumulation by bacteria. Comp Biochem Physiol A Mol Integr Physiol, 2001; 130:437–460.
70.
ZaytsevaE, ErmolaevaS, SomovGP. Low genetic diversity and epidemiological significance of Listeria monocytogenes isolated from wild animals in the far east of Russia. Infect Genet Evol, 2007; 7:736–742.
71.
ZhangZ, SchwartzS, WagnerL, MillerW. A greedy algorithm for aligning DNA sequences. J Comput Biol, 2000; 7:203–214.
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