Gene Expression Analysis of Salmonella enterica Enteritidis Nal R and Salmonella enterica Kentucky 3795 Exposed to HCl and Acetic Acid in Rich Medium

Abstract

In the United States, serovar Kentucky has become one of the most frequently isolated Salmonella enterica serovars from chickens. The reasons for this prevalence are not well understood. Phenotypic comparisons of poultry Salmonella isolates belonging to various serovars demonstrated that serovar Kentucky isolates differed from those of most other serovars in their response to acid. Microarray and qPCR analyses were performed with aerated exponentially growing poultry isolates, Salmonella enterica serovar Kentucky 3795 and Enteritidis Nal^R, exposed for 10 min to tryptic soy broth (TSB) adjusted to pH 4.5 with HCl and to pH 5.5 with HCl or acetic acid. Data obtained by microarray analysis indicated that more genes were up- or down-regulated in strain Kentucky 3795 than in Enteritidis Nal^R under acidic conditions. Acid exposure in general caused up-regulation of energy metabolism genes and down-regulation of protein synthesis genes, particularly of ribosomal protein genes. Both strains appear to similarly utilize the lysine-based pH homeostasis system, as up-regulation of cadB was observed under the acidic conditions. Expression of regulatory genes (rpoS, fur, phoPQ) known to be involved in the acid response showed similar trends in both isolates. Differences between Kentucky 3795 and Enteritidis Nal^R were observed with respect to the expression of the hdeB-like locus SEN1493 (potentially encoding a chaperone important to acid response), and some differences in the expression of other genes such as those involved in citrate utilization and motility were noted. It appears that the early stages of the transcriptional response to acid by isolates Kentucky 3795 and Enteritidis Nal^R are similar, but differences exist in the scope and in some facets of the response. Possibly, the quantitative differences observed might lead to differences in protein levels that could explain the observed differences in the acid phenotype of serovar Kentucky and other Salmonella serovars.

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References

Achard

MES

, Tree

, Holden

et al. The multi-copper-ion oxidase CueO of Salmonella enterica serovar Typhimurium is required for systemic virulence. Infect Immun, 2010; 78:2312–2319.

Adams

, Fowler

, Kinsella

et al. Proteomic detection of PhoPQ- and acid-mediated repression of Salmonella motility. Proteomics, 2001; 1:597–607.

Altschul

, Madden

, Schäffer

et al. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res, 1997; 25:3389–3402.

Berrang

, Bailey

, Altekruse

et al. Prevalence, serotype, and antimicrobial resistance of Salmonella on broiler carcasses postpick and postchill in 20 US processing plants. J Food Prot, 2009; 72:1610–1615.

, Galan

. Selective inhibition of Type III secretion activated signaling by the Salmonella effector AvrA. PLOS Pathogens, 2009; 9:e1000595.

Espariz

, Checa

, Perez Audero

et al. 2007. Dissecting the Salmonella response to copper. Microbiology, 2007; 153:2989–2997.

Foster

. Low pH adaptation and the acid tolerance response of Salmonella Typhimurium. Crit Rev Microbiol, 1995; 21:215–237.

Foster

, Hall

. Adaptice acidification tolerance response of Salmonella Typhimurium. J Bacteriol, 1990; 172:771–778.

Foster

, Hall

. Inducible pH homeostasis and the acid tolerance response of Salmonella Typhimurium. J Bacteriol, 1991; 173:5129–5135.

10.

Fricke

, McDermott

, Mammel

et al. Antimicrobial resistance-conferring plasmids with similarity to virulence plasmids from avian pathogenic Escherichia coli strains in Salmonella enterica serovar Kentucky isolates from poultry. Appl Environ Microbiol, 2009; 75:5963–5971.

11.

Henry

, Couillault

, Rockenfeller

et al. The Salmonella effector protein PipB2 is a linker for kinesin-1. Proc Natl Acad Sci USA, 2006; 103:13497–13502.

12.

Hoskins

, Pak

, Maurizi

et al. The role of the ClpA chaperone in proteolysis by ClpAP. Proc Natl Acad Sci USA, 1998; 95:12135–12140.

13.

Joerger

, Sartori

, Kniel

. Comparison of genetic and physiological properties of Salmonella enterica isolates from chickens reveals one major difference between serovar Kentucky and other serovars: Response to acid. Foodborne Pathog Dis, 2009; 6:503–512.

14.

Joerger

, Hanning

, Ricke

. Presence of arsenic resistance in Salmonella enterica serovar Kentucky and other serovars isolated from poultry. Avian Dis, 2010; 54:1178–1182.

15.

Johnson

, Thorsness

, Anderson

et al. Horizontal gene transfer of a ColV plasmid has resulted in a dominant avian clonal type of Salmonella enterica serovar Kentucky. PLOS One, 2010; 5:e15524.

16.

Kehres

, Janakiraman

, Slauch

et al. SitABCD is the alkaline Mn^2+_transporter of Salmonella enterica serovar Typhimurium. J Bacteriol, 2002; 184:3159–3166.

17.

Kelly

, Goldberg

, Carroll

et al. A global role for Fis in the transcriptional control of metabolism and type III secretion in Salmonella enterica serovar Typhimurium. Microbiology, 2004; 150:2037–2053.

18.

Kern

, Malki

, Abdallah

et al. Escherichia coli HdeB is an acid stress chaperone. J Bacteriol, 2007; 189:603–610.

19.

Kieboom

, Abee

. Arginine-dependent acid resistance in Salmonella enterica serovar Typhimurium. J Bacteriol, 2006; 188:5650–5653.

20.

Lee

, Kim

et al. CadC has a global translational effect during acid adaptation in Salmonella enterica serovar Typhimurium. J Bacteriol, 2007; 189:2417–2425.

21.

Lestari

, Han

, Fei

et al. Prevalence and antimicrobial resistance of Salmonella serovars in conventional and organic chickens from Louisiana retail stores. J Food Prot, 2009; 72:1165–1172.

22.

Macomber

, Imlay

. The iron-sulfur clusters of dehydratases are primary intracellular targets of copper toxicity. Proc Natl Acad Sci USA, 2009; 106:8344–8349.

23.

Maurelli

, Fernandez

, Bloch

et al. “Black holes” and bacterial pathogenicity: A large genomic deletion that enhances the virulence of Shigella spp. and enteroinvasive Escherichia coli. Proc Natl Acad Sci USA, 1998; 95:3943–3948.

24.

Melendez

, Hanning

, Han

et al. Salmonella enterica isolates from pasture-raised poultry exhibit antimicrobial resistance and class I integrons. J Appl Microbiol, 2010; 109:1957–1966.

25.

Outten

, Wood

, Munoz

et al. The SufE protein and the SufBCD complex enhance SufS cysteine desulfurase activity as part of a sulfur transfer pathway for Fe-S cluster assembly in Escherichia coli. J Biol Chem, 2003; 278:45713–45719.

26.

Park

, Bearson

, Bang

et al. Internal pH crisis, lysine decarboxylase and the acid tolerance response of Salmonella Typhimurium. Mol Microbiol, 1996; 20:605–611.

27.

Porwollik

, Wong

RMY

, Sims

et al. The Delta uvrB mutations in the Ames strains of Salmonella span 15 to 119 genes. Mutation Res, 2001; 1-2:1–11.

28.

Porwollik

, Frye

, Florea

et al. A non-redundant microarray of genes for two related bacteria. Nucleic Acids Res, 2003; 31:1869–1876.

29.

Porwollik

, Santiviago

, Cheng

et al. Differences in gene content between Salmonella enterica serovar enteritidis isolates and comparison to closely related serovars Gallinarum and Dublin. J Bacteriol, 2005; 187:6545–6555.

30.

Riordan

, Tietjen

, Walsh

et al. Inactivation of alternative sigma factor 54 (RpoN) leads to increased acid resistance, and alters locus of enterocyte effacement (LEE) expression in Escherichia coli O157:H7. Microbiology, 2010; 156:719–730.

31.

Ruiz

, McMurry

, Levy

. Role of the multidrug resistance regulator MarA in global regulation of the hdeAB acid resistance operon in Escherichia coli. J Bacteriol, 2008; 190:1290–1297.

32.

Sauer

, Canonaco

, Heri

et al. The soluble and membrane-bound transhydrogenases UdhA and PntAB have divergent functions in NADPH metabolism of Escherichia coli. J Biol Chem, 2004; 279:6613–6619.

33.

Šeputiene

, Motiejûnas

, Sužiedelis

et al. Molecular characterization of the acid-inducible asr gene of Escherichia coli and its role in acid stress response. J Bacteriol, 2003; 185:2475–2484.

34.

Seputiene

, Suziedelis

, Normark

et al. Transcriptional analysis of the acid-inducible asr gene in enterobacteria. Res Microbiol, 2004; 155:535–542.

35.

Suziedeliene

, Suziedelis

, Garbenciute

et al. The acid-inducible asr gene in Escherichia coli: Transcriptional control by the phoBR operon. J Bacteriol, 1999; 181:2084–2093.

36.

Ueta

, Yoshida

, Wada

et al. Ribosome binding proteins YhbH and YfiA have opposite functions during 100S formation in the stationary phase of Escherichia coli. Genes Cells, 2005; 10:1103–1112.

37.

Untergasser

, Nijveen

, Rao

et al. Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res, 2007; 35:W71–W74.

38.

[USDA] United States Department of Agriculture–-Food Safety Inspection Services (FSIS). Serotypes profile of Salmonella Isolates from meat and poultry products, January 1998 through December 2007. http://www.fsis.usda.gov/Science/Serotypes_Profile_Salmonella_Isolates/index.asp. 2011 December 15.

39.

Vazquez-Juarez

, Kuriakose

, Rasko

et al. CadA negatively regulates Escherichia coli O157:H7 adherence and intestinal colonization. Infect Immun, 2008; 76:5072–5081.

40.

Wang

, Phillippy

, Deng

et al. Transcriptomic response of Salmonella enterica serovars Enteritidis and Typhimurium to chlorine-based oxidative stress. Appl Environ Microbiol, 2010; 76:5013–5024.

41.

Xia

, McClelland

, Wang

. WebArray: An online platform for microarray data analysis. BMC Bioinform, 2005; 6:306.

42.

Xia

, McClelland

, Porwollik

et al. WebArrayDB: Cross-platform microarray data analysis and public data repository. Bioinformatics, 2009; 25:2425–2429.

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