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The small RNA Aar in Acinetobacter baylyi: a putative regulator of amino acid metabolism

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Abstract

Small non-coding RNAs (sRNAs) are key players in prokaryotic metabolic circuits, allowing the cell to adapt to changing environmental conditions. Regulatory interference by sRNAs in cellular metabolism is often facilitated by the Sm-like protein Hfq. A search for novel sRNAs in A. baylyi intergenic regions was performed by a biocomputational screening. One candidate, Aar, encoded between trpS and sucD showed Hfq dependency in Northern blot analysis. Aar was expressed strongly during stationary growth phase in minimal medium; in contrast, in complex medium, strongest expression was in the exponential growth phase. Whereas over-expression of Aar in trans did not affect bacterial growth, seven mRNA targets predicted by two in silico approaches were upregulated in stationary growth phase. All seven mRNAs are involved in A. baylyi amino acid metabolism. A putative binding site for Lrp, the global regulator of branched-chain amino acids in E. coli, was observed within the aar gene. Both facts imply an Aar participation in amino acid metabolism.

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References

  • Altuvia S (2007) Identification of bacterial small non-coding RNAs: experimental approaches. Curr Opin Microbiol 10:257–261

    Article  PubMed  CAS  Google Scholar 

  • Argaman L, Hershberg R, Vogel J, Bejerano G, Wagner EG, Margalit H, Altuvia S (2001) Novel small RNA-encoding genes in the intergenic regions of Escherichia coli. Curr Biol 11:941–950

    Article  PubMed  CAS  Google Scholar 

  • Axmann IM, Kensche P, Vogel J, Kohl S, Herzel H, Hess WR (2005) Identification of cyanobacterial non-coding RNAs by comparative genome analysis. Genome Biol 6:R73

    Article  PubMed  CAS  Google Scholar 

  • Barbe V, Vallenet D, Fonknechten N, Kreimeyer A, Oztas S, Labarre L, Cruveiller S, Robert C, Duprat S, Wincker P, Ornston LN, Weissenbach J, Marliere P, Cohen GN, Medigue C (2004) Unique features revealed by the genome sequence of Acinetobacter sp. ADP1, a versatile and naturally transformation competent bacterium. Nucleic Acids Res 32:5766–5779

    Article  PubMed  CAS  Google Scholar 

  • Bejerano-Sagie M, Xavier KB (2007) The role of small RNAs in quorum sensing. Curr Opin Microbiol 10:189–198

    Article  PubMed  CAS  Google Scholar 

  • Boisset S, Geissmann T, Huntzinger E, Fechter P, Bendridi N, Possedko M, Chevalier C, Helfer AC, Benito Y, Jacquier A, Gaspin C, Vandenesch F, Romby P (2007) Staphylococcus aureus RNAIII coordinately represses the synthesis of virulence factors and the transcription regulator Rot by an antisense mechanism. Genes Dev 21:1353–1366

    Article  PubMed  CAS  Google Scholar 

  • Brescia CC, Mikulecky PJ, Feig AL, Sledjeski DD (2003) Identification of the Hfq-binding site on DsrA RNA: Hfq binds without altering DsrA secondary structure. Rna 9:33–43

    Article  PubMed  CAS  Google Scholar 

  • Buck D, Guest JR (1989) Overexpression and site-directed mutagenesis of the succinyl-CoA synthetase of Escherichia coli and nucleotide sequence of a gene (g30) that is adjacent to the suc operon. Biochem J 260:737–747

    PubMed  CAS  Google Scholar 

  • Busch A, Richter AS, Backofen R (2008) IntaRNA: efficient prediction of bacterial sRNA targets incorporating target site accessibility and seed regions. Bioinformatics 24:2849–2856

    Article  PubMed  CAS  Google Scholar 

  • Calvo JM, Matthews RG (1994) The leucine-responsive regulatory protein, a global regulator of metabolism in Escherichia coli. Microbiol Rev 58:466–490

    PubMed  CAS  Google Scholar 

  • Cui Y, Wang Q, Stormo GD, Calvo JM (1995) A consensus sequence for binding of Lrp to DNA. J Bacteriol 177:4872–4880

    PubMed  CAS  Google Scholar 

  • de Berardinis V, Vallenet D, Castelli V, Besnard M, Pinet A, Cruaud C, Samair S, Lechaplais C, Gyapay G, Richez C, Durot M, Kreimeyer A, Le Fevre F, Schachter V, Pezo V, Doring V, Scarpelli C, Medigue C, Cohen GN, Marliere P, Salanoubat M, Weissenbach J (2008) A complete collection of single-gene deletion mutants of Acinetobacter baylyi ADP1. Mol Syst Biol 4:174

    PubMed  Google Scholar 

  • de Boer PA, Crossley RE, Rothfield LI (1989) A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli. Cell 56:641–649

    PubMed  Google Scholar 

  • Delihas N, Forst S (2001) MicF: an antisense RNA gene involved in response of Escherichia coli to global stress factors. J Mol Biol 313:1–12

    Article  PubMed  CAS  Google Scholar 

  • Figurski DH, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

    Article  PubMed  CAS  Google Scholar 

  • Folichon M, Allemand F, Regnier P, Hajnsdorf E (2005) Stimulation of poly(A) synthesis by Escherichia coli poly(A)polymerase I is correlated with Hfq binding to poly(A) tails. FEBS J 272:454–463

    Article  PubMed  CAS  Google Scholar 

  • Forner J, Weber B, Thuss S, Wildum S, Binder S (2007) Mapping of mitochondrial mRNA termini in Arabidopsis thaliana: t-elements contribute to 5′ and 3′ end formation. Nucleic Acids Res 35:3676–3692

    Article  PubMed  CAS  Google Scholar 

  • Gardner PP, Daub J, Tate JG, Nawrocki EP, Kolbe DL, Lindgreen S, Wilkinson AC, Finn RD, Griffiths-Jones S, Eddy SR, Bateman A (2009) Rfam: updates to the RNA families database. Nucleic Acids Res 37:D136–D140

    Article  PubMed  CAS  Google Scholar 

  • Geissmann TA, Touati D (2004) Hfq, a new chaperoning role: binding to messenger RNA determines access for small RNA regulator. EMBO J 23:396–405

    Article  PubMed  CAS  Google Scholar 

  • Gorke B, Vogel J (2008) Non-coding RNA control of the making and breaking of sugars. Genes Dev 22:2914–2925

    Article  PubMed  CAS  Google Scholar 

  • Gottesman S (2005) Micros for microbes: non-coding regulatory RNAs in bacteria. Trends Genet 21:399–404

    Article  PubMed  CAS  Google Scholar 

  • Hajnsdorf E, Regnier P (2000) Host factor Hfq of Escherichia coli stimulates elongation of poly(A) tails by poly(A) polymerase I. Proc Natl Acad Sci USA 97:1501–1505

    Article  PubMed  CAS  Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Article  PubMed  CAS  Google Scholar 

  • Hershberg R, Altuvia S, Margalit H (2003) A survey of small RNA-encoding genes in Escherichia coli. Nucleic Acids Res 31:1813–1820

    Article  PubMed  CAS  Google Scholar 

  • Kalamorz F, Reichenbach B, Marz W, Rak B, Gorke B (2007) Feedback control of glucosamine-6-phosphate synthase GlmS expression depends on the small RNA GlmZ and involves the novel protein YhbJ in Escherichia coli. Mol Microbiol 65:1518–1533

    Article  PubMed  CAS  Google Scholar 

  • Kawamoto H, Koide Y, Morita T, Aiba H (2006) Base-pairing requirement for RNA silencing by a bacterial small RNA and acceleration of duplex formation by Hfq. Mol Microbiol 61:1013–1022

    Article  PubMed  CAS  Google Scholar 

  • Kawano M, Reynolds AA, Miranda-Rios J, Storz G (2005) Detection of 5′- and 3′-UTR-derived small RNAs and cis-encoded antisense RNAs in Escherichia coli. Nucleic Acids Res 33:1040–1050

    Article  PubMed  CAS  Google Scholar 

  • Keen NT, Tamaki S, Kobayashi D, Trollinger D (1988) Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene 70:191–197

    Article  PubMed  CAS  Google Scholar 

  • Kingsford CL, Ayanbule K, Salzberg SL (2007) Rapid, accurate, computational discovery of Rho-independent transcription terminators illuminates their relationship to DNA uptake. Genome Biol 8:R22

    Article  PubMed  CAS  Google Scholar 

  • Le Derout J, Folichon M, Briani F, Deho G, Regnier P, Hajnsdorf E (2003) Hfq affects the length and the frequency of short oligo(A) tails at the 3′ end of Escherichia coli rpsO mRNAs. Nucleic Acids Res 31:4017–4023

    Article  PubMed  CAS  Google Scholar 

  • Le Derout J, Boni IV, Regnier P, Hajnsdorf E (2010) Hfq affects mRNA levels independently of degradation. BMC Mol Biol 11:17

    Article  PubMed  CAS  Google Scholar 

  • Lin R, D’Ari R, Newman EB (1992) Lambda placMu insertions in genes of the leucine regulon: extension of the regulon to genes not regulated by leucine. J Bacteriol 174:1948–1955

    PubMed  CAS  Google Scholar 

  • Liu MY, Gui G, Wei B, Preston JF 3rd, Oakford L, Yuksel U, Giedroc DP, Romeo T (1997) The RNA molecule CsrB binds to the global regulatory protein CsrA and antagonizes its activity in Escherichia coli. J Biol Chem 272:17502–17510

    Article  PubMed  CAS  Google Scholar 

  • Livny J, Waldor MK (2007) Identification of small RNAs in diverse bacterial species. Curr Opin Microbiol 10:96–101

    Article  PubMed  CAS  Google Scholar 

  • Livny J, Brencic A, Lory S, Waldor MK (2006) Identification of 17 Pseudomonas aeruginosa sRNAs and prediction of sRNA-encoding genes in 10 diverse pathogens using the bioinformatic tool sRNAPredict2. Nucleic Acids Res 34:3484–3493

    Article  PubMed  CAS  Google Scholar 

  • Majdalani N, Cunning C, Sledjeski D, Elliott T, Gottesman S (1998) DsrA RNA regulates translation of RpoS message by an anti-antisense mechanism, independent of its action as an antisilencer of transcription. Proc Natl Acad Sci USA 95:12462–12467

    Article  PubMed  CAS  Google Scholar 

  • Maki K, Uno K, Morita T, Aiba H (2008) RNA, but not protein partners, is directly responsible for translational silencing by a bacterial Hfq-binding small RNA. Proc Natl Acad Sci USA 105:10332–10337

    Article  PubMed  Google Scholar 

  • Masse E, Gottesman S (2002) A small RNA regulates the expression of genes involved in iron metabolism in Escherichia coli. Proc Natl Acad Sci USA 99:4620–4625

    Article  PubMed  CAS  Google Scholar 

  • Masse E, Escorcia FE, Gottesman S (2003) Coupled degradation of a small regulatory RNA and its mRNA targets in Escherichia coli. Genes Dev 17:2374–2383

    Article  PubMed  CAS  Google Scholar 

  • Masse E, Salvail H, Desnoyers G, Arguin M (2007) Small RNAs controlling iron metabolism. Curr Opin Microbiol 10:140–145

    Article  PubMed  CAS  Google Scholar 

  • Mizuno T, Chou MY, Inouye M (1984) A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci USA 81:1966–1970

    Article  PubMed  CAS  Google Scholar 

  • Moll I, Afonyushkin T, Vytvytska O, Kaberdin VR, Blasi U (2003) Coincident Hfq binding and RNase E cleavage sites on mRNA and small regulatory RNAs. Rna 9:1308–1314

    Article  PubMed  CAS  Google Scholar 

  • Mückstein U, Tafer H, Hackermuller J, Bernhart SH, Stadler PF, Hofacker IL (2006) Thermodynamics of RNA-RNA binding. Bioinformatics 22:1177–1182

    Article  PubMed  CAS  Google Scholar 

  • Munch R, Hiller K, Barg H, Heldt D, Linz S, Wingender E, Jahn D (2003) PRODORIC: prokaryotic database of gene regulation. Nucleic Acids Res 31:266–269

    Article  PubMed  CAS  Google Scholar 

  • Newman EB, Lin R (1995) Leucine-responsive regulatory protein: a global regulator of gene expression in E. coli. Annu Rev Microbiol 49:747–775

    Article  PubMed  CAS  Google Scholar 

  • Oelmüller U, Krüger N, Steinbüchel A, Freidrich CG (1990) Isolation of prokaryotic RNA and detection of specific mRNA with biotinylated probes. J Microbiol Methods 11:73–81

    Article  Google Scholar 

  • Pfeiffer V, Sittka A, Tomer R, Tedin K, Brinkmann V, Vogel J (2007) A small non-coding RNA of the invasion gene island (SPI-1) represses outer membrane protein synthesis from the Salmonella core genome. Mol Microbiol 66:1174–1191

    Article  PubMed  CAS  Google Scholar 

  • Regnier P, Hajnsdorf E (2008) The role of RNA chaperone Hfq in poly(A) metabolism methods to determine positions, abundance, and lengths of short oligo(A) tails. Methods Enzymol 447:161–181

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Schilling D, Gerischer U (2009) The Acinetobacter baylyi hfq gene encodes a large protein with an unusual C terminus. J Bacteriol 191:5553–5562

    Article  PubMed  CAS  Google Scholar 

  • Schimmel PR, Soll D (1979) Aminoacyl-tRNA synthetases: general features and recognition of transfer RNAs. Annu Rev Biochem 48:601–648

    Article  PubMed  CAS  Google Scholar 

  • Sharma CM, Darfeuille F, Plantinga TH, Vogel J (2007) A small RNA regulates multiple ABC transporter mRNAs by targeting C/A-rich elements inside and upstream of ribosome-binding sites. Genes Dev 21:2804–2817

    Article  PubMed  CAS  Google Scholar 

  • Sonnleitner E, Sorger-Domenigg T, Madej MJ, Findeiss S, Hackermuller J, Huttenhofer A, Stadler PF, Blasi U, Moll I (2008) Detection of small RNAs in Pseudomonas aeruginosa by RNomics and structure-based bioinformatic tools. Microbiology 154:3175–3187

    Article  PubMed  CAS  Google Scholar 

  • Soper TJ, Woodson SA (2008) The rpoS mRNA leader recruits Hfq to facilitate annealing with DsrA sRNA. Rna 14:1907–1917

    Article  PubMed  CAS  Google Scholar 

  • Starmer J, Stomp A, Vouk M, Bitzer D (2006) Predicting Shine-Dalgarno sequence locations exposes genome annotation errors. PLoS Comput Biol 2:e57

    Article  PubMed  CAS  Google Scholar 

  • Trautwein G, Gerischer U (2001) Effects exerted by transcriptional regulator PcaU from Acinetobacter sp. strain ADP1. J Bacteriol 183:873–881

    Article  PubMed  CAS  Google Scholar 

  • Tu KC, Bassler BL (2007) Multiple small RNAs act additively to integrate sensory information and control quorum sensing in Vibrio harveyi. Genes Dev 21:221–233

    Article  PubMed  CAS  Google Scholar 

  • Udekwu KI, Darfeuille F, Vogel J, Reimegard J, Holmqvist E, Wagner EG (2005) Hfq-dependent regulation of OmpA synthesis is mediated by an antisense RNA. Genes Dev 19:2355–2366

    Article  PubMed  CAS  Google Scholar 

  • Valentin-Hansen P, Eriksen M, Udesen C (2004) The bacterial Sm-like protein Hfq: a key player in RNA transactions. Mol Microbiol 51:1525–1533

    Article  PubMed  CAS  Google Scholar 

  • Vanderpool CK (2007) Physiological consequences of small RNA-mediated regulation of glucose-phosphate stress. Curr Opin Microbiol 10:146–151

    Article  PubMed  CAS  Google Scholar 

  • Vaneechoutte M, Young DM, Ornston LN, De Baere T, Nemec A, Van Der Reijden T, Carr E, Tjernberg I, Dijkshoorn L (2006) Naturally transformable Acinetobacter sp. strain ADP1 belongs to the newly described species Acinetobacter baylyi. Appl Environ Microbiol 72:932–936

    Article  PubMed  CAS  Google Scholar 

  • Vogel J, Papenfort K (2006) Small non-coding RNAs and the bacterial outer membrane. Curr Opin Microbiol 9:605–611

    Article  PubMed  CAS  Google Scholar 

  • Voss B, Georg J, Schon V, Ude S, Hess WR (2009) Biocomputational prediction of non-coding RNAs in model cyanobacteria. BMC Genomics 10:123

    Article  PubMed  CAS  Google Scholar 

  • Washietl S, Hofacker IL, Stadler PF (2005) Fast and reliable prediction of non-coding RNAs. Proc Natl Acad Sci USA 102:2454–2459

    Article  PubMed  CAS  Google Scholar 

  • Wassarman KM (2007) 6S RNA: a small RNA regulator of transcription. Curr Opin Microbiol 10:164–168

    Article  PubMed  CAS  Google Scholar 

  • Willkomm DK, Minnerup J, Hüttenhofer A, Hartmann RK (2005) Experimental RNomics in Aquifex aeolicus: identification of small non-coding RNAs and the putative 6S RNA homolog. Nucleic Acids Res 33:1949–1960

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We would like to thank Björn Voss for his predictions of sRNA genes in Acinetobacter and Véronique de Berardinis for her effort to delete aar. Furthermore, we would like to thank Iris Steiner for her contribution. This work was supported by the German Federal Ministry of Education and Research (BMBF grant 0313921 FRISYS to A.S.R.); the German Research Foundation (DFG grant SPP1258 to S.F. STA850/7-1 and A.S.R. BA2168/2-1); and the state of Baden-Württemberg, Germany (personal LGFG grant to D.S.).

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Correspondence to Dominik Schilling.

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Communicated by Wolfgang Buckel.

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Schilling, D., Findeiß, S., Richter, A.S. et al. The small RNA Aar in Acinetobacter baylyi: a putative regulator of amino acid metabolism. Arch Microbiol 192, 691–702 (2010). https://doi.org/10.1007/s00203-010-0592-6

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  • DOI: https://doi.org/10.1007/s00203-010-0592-6

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