All Title Author
Keywords Abstract


Intrinsic Thermal Sensing Controls Proteolysis of Yersinia Virulence Regulator RovA

DOI: 10.1371/journal.ppat.1000435

Full-Text   Cite this paper   Add to My Lib

Abstract:

Pathogens, which alternate between environmental reservoirs and a mammalian host, frequently use thermal sensing devices to adjust virulence gene expression. Here, we identify the Yersinia virulence regulator RovA as a protein thermometer. Thermal shifts encountered upon host entry lead to a reversible conformational change of the autoactivator, which reduces its DNA-binding functions and renders it more susceptible for proteolysis. Cooperative binding of RovA to its target promoters is significantly reduced at 37°C, indicating that temperature control of rovA transcription is primarily based on the autoregulatory loop. Thermally induced reduction of DNA-binding is accompanied by an enhanced degradation of RovA, primarily by the Lon protease. This process is also subject to growth phase control. Studies with modified/chimeric RovA proteins indicate that amino acid residues in the vicinity of the central DNA-binding domain are important for proteolytic susceptibility. Our results establish RovA as an intrinsic temperature-sensing protein in which thermally induced conformational changes interfere with DNA-binding capacity, and secondarily render RovA susceptible to proteolytic degradation.

References

[1]  Konkel ME, Tilly K (2000) Temperature-regulated expression of bacterial virulence genes. Microbes Infect 2: 157–166.
[2]  Schumann W (2007) Thermosensors in eubacteria: role and evolution. J Biosci 32: 549–557.
[3]  Straley SC, Perry RD (1995) Environmental modulation of gene expression and pathogenesis in Yersinia. Trends Microbiol 3: 310–317.
[4]  Stenseth NC, Atshabar BB, Begon M, Belmain SR, Bertherat E, et al. (2008) Plague: past, present, and future. PLoS Med 5: e3. doi:10.1371/journal.pmed.0050003.
[5]  Bottone EJ (1997) Yersinia enterocolitica: the charisma continues. Clin Microbiol Rev 10: 257–276.
[6]  Koornhof HJ, Smego RA Jr, Nicol M (1999) Yersiniosis. II: The pathogenesis of Yersinia infections. Eur J Clin Microbiol Infect Dis 18: 87–112.
[7]  Marceau M (2005) Transcriptional regulation in Yersinia: an update. Curr Issues Mol Biol 7: 151–177.
[8]  Motin VL, Georgescu AM, Fitch JP, Gu PP, Nelson DO, et al. (2004) Temporal global changes in gene expression during temperature transition in Yersinia pestis. J Bacteriol 186: 6298–6305.
[9]  Bolin I, Norlander I, Wolf-Watz H (1982) Temperature-inducible outer membrane protein of Yersinia pseudotuberculosis and Yersinia enterocolitica is associated with the virulence plasmid. Infect Immun 37: 506–512.
[10]  Rohde JR, Luan XS, Rohde H, Fox JM, Minnich SA (1999) The Yersinia enterocolitica pYV virulence plasmid contains multiple intrinsic DNA bends which melt at 37 degrees C. J Bacteriol 181: 4198–4204.
[11]  Jackson MW, Silva-Herzog E, Plano GV (2004) The ATP-dependent ClpXP and Lon proteases regulate expression of the Yersinia pestis type III secretion system via regulated proteolysis of YmoA, a small histone-like protein. Mol Microbiol 54: 1364–1378.
[12]  Hoe NP, Goguen JD (1993) Temperature sensing in Yersinia pestis: translation of the LcrF activator protein is thermally regulated. J Bacteriol 175: 7901–7909.
[13]  Thomson NR, Cox A, Bycroft BW, Stewart GS, Williams P, et al. (1997) The Rap and Hor proteins of Erwinia, Serratia and Yersinia: a novel subgroup in a growing superfamily of proteins regulating diverse physiological processes in bacterial pathogens. Mol Microbiol 26: 531–544.
[14]  Ellison DW, Miller VL (2006) Regulation of virulence by members of the MarR/SlyA family. Curr Opin Microbiol 9: 153–159.
[15]  Heroven AK, B?hme K, Tran-Winkler H, Dersch P (2007) Regulatory elements implicated in the environmental control of invasin expression in enteropathogenic Yersinia. Adv Exp Med Biol 603: 156–166.
[16]  Revell PA, Miller VL (2000) A chromosomally encoded regulator is required for expression of the Yersinia enterocolitica inv gene and for virulence. Mol Microbiol 35: 677–685.
[17]  Nagel G, Lahrz A, Dersch P (2001) Environmental control of invasin expression in Yersinia pseudotuberculosis is mediated by regulation of RovA, a transcriptional activator of the SlyA/Hor family. Mol Microbiol 41: 1249–1269.
[18]  Heroven AK, Dersch P (2006) RovM, a novel LysR-type regulator of the virulence activator gene rovA, controls cell invasion, virulence and motility of Yersinia pseudotuberculosis. Mol Microbiol 62: 1469–1483.
[19]  Cathelyn JS, Crosby SD, Lathem WW, Goldman WE, Miller VL (2006) RovA, a global regulator of Yersinia pestis, specifically required for bubonic plague. Proc Natl Acad Sci U S A 103: 13514–13519.
[20]  Heroven AK, Nagel G, Tran HJ, Parr S, Dersch P (2004) RovA is autoregulated and antagonizes H-NS-mediated silencing of invasin and rovA expression in Yersinia pseudotuberculosis. Mol Microbiol 53: 871–888.
[21]  Lawrenz MB, Miller VL (2007) Comparative analysis of the regulation of rovA from the pathogenic yersiniae. J Bacteriol 189: 5963–5975.
[22]  Ellison DW, Miller VL (2006) H-NS represses inv transcription in Yersinia enterocolitica through competition with RovA and interaction with YmoA. J Bacteriol 188: 5101–5112.
[23]  Tran HJ, Heroven AK, Winkler L, Spreter T, Beatrix B, et al. (2005) Analysis of RovA, a transcriptional regulator of Yersinia pseudotuberculosis virulence that acts through antirepression and direct transcriptional activation. J Biol Chem 280: 42423–42432.
[24]  Heroven AK, B?hme K, Rohde M, Dersch P (2008) A Csr-type regulatory system, including small non-coding RNAs, regulates the global virulence regulator RovA of Yersinia pseudotuberculosis through RovM. Mol Microbiol 68: 1179–1195.
[25]  Lupas A (1996) Coiled coils: new structures and new functions. Trends Biochem Sci 21: 375–382.
[26]  Parsell DA, Silber KR, Sauer RT (1990) Carboxy-terminal determinants of intracellular protein degradation. Genes Dev 4: 277–286.
[27]  Keiler KC, Waller PR, Sauer RT (1996) Role of a peptide tagging system in degradation of proteins synthesized from damaged messenger RNA. Science 271: 990–993.
[28]  Wang L, Wilson S, Elliott T (1999) A mutant HemA protein with positive charge close to the N terminus is stabilized against heme-regulated proteolysis in Salmonella typhimurium. J Bacteriol 181: 6033–6041.
[29]  Wang KH, Oakes ES, Sauer RT, Baker TA (2008) Tuning the strength of a bacterial N-end rule degradation signal. J Biol Chem 283: 24600–24607.
[30]  Gonzalez M, Frank EG, Levine AS, Woodgate R (1998) Lon-mediated proteolysis of the Escherichia coli UmuD mutagenesis protein: in vitro degradation and identification of residues required for proteolysis. Genes Dev 12: 3889–3899.
[31]  Lee I, Suzuki CK (2008) Functional mechanics of the ATP-dependent Lon protease - lessons from endogenous protein and synthetic peptide substrates. Biochim Biophys Acta 1784: 727–735.
[32]  Phillips TA, VanBogelen RA, Neidhardt FC (1984) lon gene product of Escherichia coli is a heat-shock protein. J Bacteriol 159: 283–287.
[33]  Goldberg AL (1992) The mechanism and functions of ATP-dependent proteases in bacterial and animal cells. Eur J Biochem 203: 9–23.
[34]  Gal-Mor O, Valdez Y, Finlay BB (2006) The temperature-sensing protein TlpA is repressed by PhoP and dispensable for virulence of Salmonella enterica serovar Typhimurium in mice. Microbes Infect 8: 2154–2162.
[35]  Hurme R, Berndt KD, Normark SJ, Rhen M (1997) A proteinaceous gene regulatory thermometer in Salmonella. Cell 90: 55–64.
[36]  Servant P, Grandvalet C, Mazodier P (2000) The RheA repressor is the thermosensor of the HSP18 heat shock response in Streptomyces albus. Proc Natl Acad Sci U S A 97: 3538–3543.
[37]  Gottesman S (2003) Proteolysis in bacterial regulatory circuits. Annu Rev Cell Dev Biol 19: 565–587.
[38]  Licht S, Lee I (2008) Resolving individual steps in the operation of ATP-dependent proteolytic molecular machines: from conformational changes to substrate translocation and processivity. Biochemistry 47: 3595–3605.
[39]  Sauer RT, Bolon DN, Burton BM, Burton RE, Flynn JM, et al. (2004) Sculpting the proteome with AAA(+) proteases and disassembly machines. Cell 119: 9–18.
[40]  Tsilibaris V, Maenhaut-Michel G, Van Melderen L (2006) Biological roles of the Lon ATP-dependent protease. Res Microbiol 157: 701–713.
[41]  Wilkinson SP, Grove A (2006) Ligand-responsive transcriptional regulation by members of the MarR family of winged helix proteins. Curr Issues Mol Biol 8: 51–62.
[42]  Zhao G, Weatherspoon N, Kong W, Curtiss R 3rd, Shi Y (2008) A dual-signal regulatory circuit activates transcription of a set of divergent operons in Salmonella typhimurium. Proc Natl Acad Sci U S A 105: 20924–20929.
[43]  Baker TA, Sauer RT (2006) ATP-dependent proteases of bacteria: recognition logic and operating principles. Trends Biochem Sci 31: 647–653.
[44]  Gur E, Sauer RT (2008) Recognition of misfolded proteins by Lon, a AAA+ protease. Genes Dev 22: 2267–2277.
[45]  Shah IM, Wolf RE Jr (2006) Sequence requirements for Lon-dependent degradation of the Escherichia coli transcription activator SoxS: identification of the SoxS residues critical to proteolysis and specific inhibition of in vitro degradation by a peptide comprised of the N-terminal 21 amino acid residues. J Mol Biol 357: 718–731.
[46]  Shah IM, Wolf RE Jr (2006) Inhibition of Lon-dependent degradation of the Escherichia coli transcription activator SoxS by interaction with ‘soxbox’ DNA or RNA polymerase. Mol Microbiol 60: 199–208.
[47]  Legault P, Li J, Mogridge J, Kay LE, Greenblatt J (1998) NMR structure of the bacteriophage lambda N peptide/boxB RNA complex: recognition of a GNRA fold by an arginine-rich motif. Cell 93: 289–299.
[48]  Huang HC, Sherman MY, Kandror O, Goldberg AL (2001) The molecular chaperone DnaJ is required for the degradation of a soluble abnormal protein in Escherichia coli. J Biol Chem 276: 3920–3928.
[49]  Flynn JM, Levchenko I, Sauer RT, Baker TA (2004) Modulating substrate choice: the SspB adaptor delivers a regulator of the extracytoplasmic-stress response to the AAA+ protease ClpXP for degradation. Genes Dev 18: 2292–2301.
[50]  Rudyak SG, Shrader TE (2000) Polypeptide stimulators of the Ms-Lon protease. Protein Sci 9: 1810–1817.
[51]  Nomura K, Kato J, Takiguchi N, Ohtake H, Kuroda A (2004) Effects of inorganic polyphosphate on the proteolytic and DNA-binding activities of Lon in Escherichia coli. J Biol Chem 279: 34406–34410.
[52]  Pruteanu M, Neher SB, Baker TA (2007) Ligand-controlled proteolysis of the Escherichia coli transcriptional regulator ZntR. J Bacteriol 189: 3017–3025.
[53]  Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97: 6640–6645.

Full-Text

comments powered by Disqus