The expression of the Acinetobacter calcoaceticus recA gene increases in response to DNA damage independently of RecA and of development of competence for natural transformation
Author for correspondence: Klaas J. Hellingwerf. Tel: +31 20 5257055. Fax: +31 20 5257056. e-mail: A417hell@horus.sara.nl
‡ Present address: CHU Rangueil, Laboratoire de Bactériologie-Virologie-Hygiène (Bat L1), Av. Jean Poulhs, 31054 Toulouse Cedex, France.
† Present address: Division of Industrial Microbiology, Wageningen Agricultural University, Department of Food Science, Bomenweg 2, 6703 HD Wageningen, The Netherlands.
Using the lacZ operon fusion technique, the transcriptional control of the Acinetobacter calcoaceticus recA gene was studied. A low (approximately twofold) inductive capacity was observed for compounds that damage DNA and/or inhibit DNA replication, e.g. methyl methanesulfonate, mitomycin C, UV light and nalidixic acid. Induction of the recA gene by DNA damage was independent of functional RecA. The presence of the recA promoter region on a multicopy plasmid had the same effect on recA transcription as the presence of DNA-damaging agents. Thus, recA expression in A. calcoaceticus appears to be regulated in a novel fashion, possibly involving a non-LexA-like repressor. Regulation of the recA gene in A. calcoaceticus appears not to be part of a regulon responsible for competence for natural transformation: in cells exhibiting extremely low transformation frequencies, the level of transcription of the recA gene was found to be comparable to the level found in cells in the state of maximal competence.
BagdasarianM., LurzR., RückertB., FranklinF.C.H., BagdasarianM.M., FreyJ., TimmisK.N. Specific-purpose plasmid cloning vectors II Broad host range, high copy number, RSFlOlO-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene1981; 16:237–247
BradfordM.M. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem1976; 72:248–254
CheoD.L., BaylesK.W., YasbinR.E. Cloning and characterization of DNA damage-inducible promoter regions from Bacillus subtilis. J Bacteriol1991; 173:1696–1703
CheoD.L., BaylesK.W., YasbinR.E. Elucidation of regulatory elements that control damage induction and competence induction of the Bacillus subtilis SOS system. J Bacteriol1993; 175:5907–5915
De VosW.M., VenemaG. Transformation of Bacillus subtilis competent cells: identification of a protein involved in recombination. Mol & Gen Genet1982; 187:439–445
Fernandez De HenestrosaA., CaleroA.R., BarbéJ. Expression of the recA gene of Escherichia coli in several species of gram-negative bacteria. Mol & Gen Genet1991; 226:503–506
Gregg-JollyL.A., OrnstonL.N. Properties of Acinetohacter calcoaceticus recA and its contribution to intracellular gene conversion. Mol Microbiol1994; 12:985–992
GuerryP., PopeP.M., BurrD.H., LeiterJ., JosephS., W. & BourgeoisA.L. Development and characterization of recA mutants of Campylobacter jejuni for inclusion in attenuated vaccines. Inject Immun1994; 62:426–432
HigashitaniN., HigashitaniA., RothA., HoriuchiK. SOS induction in Escherichia coli by infection with mutant filamentous phage that are defective in initiation of complementary-strand DNA synthesis. J Bacteriol1992; 174:1612–1618
HornJ.M., OhmanD.E. Autogenous regulation and kinetics of induction of Pseudomonas aeruginosa recA transcription as analyzed with operon fusions. J Bacteriol1988; 170:4699–4705
HungerM., SchmuckerR., KishanV., HillenW. Analysis and nucleotide sequence of an origin of DNA replication in Acinetohacter calcoaceticus and its use for Escherichia coli shuttle plasmids. Gene1990; 87:45–51
KokR.G., ChristoffelsV.M., VosmanB., HellingwerfK.J. Growth-phase-dependent expression of the lipolytic system of Acinetohacter calcoaceticus BD413: cloning of a gene encoding one of the esterases. J Gen Microbiol1993; 139:2329–2342
LovettC.M., LoveP.E., YasbinR.E., 8, RobertsJ.W. SOS-like induction in Bacillus subtilis: induction of the RecA protein analog and a damage-inducible operon by DNA damage in Rec+ and DNA repair-deficient strains. J Bacteriol1988; 170:1467–1474
MartinB., GarcíaP., CastanteM.-P., ClaverysJ.P. The recA gene of Streptococcus pneumoniae is part of a competence-induced operon and controls lysogenic induction. Mol Microbiol1995; 15:367–379
MastersC.I., SmithM.D., GutmanP.D., MintonK.W. Heterozygosity and instability of amplified chromosomal insertions in the radioresistant bacterium Deinococcus radiodurans. J Bacteriol1991; 173:6110–6117
PalmenR., VosmanB., KokR., Van Der ZeeJ.R., HellingwerfK.J. Characterization of transformation-deficient mutants of Acinetohacter calcoaceticus. Mol Microbiol1992; 6:1747–1754
RamesarR.S., AbrattV., WoodsD.R., RawlingsD.E. Nucleotide sequence and expression of a cloned Thiobacillus ferrooxidans rec A gene in Escherichia coli. Gene1989; 78:1–8
Raymond-DeniseA., GuillenN. Expression of the Bacillus subtilis dinR and rec A genes after DNA damage and during competence. J Bacteriol1992; 174:3171–3176
StoryR.M., BishopD.K., KlecknerN., SteitzT.A. Structural relationship of bacterial RecA proteins to recombination proteins from bacteriophage T4 and yeast. Science1993; 259:1892–1896
VieiraJ., MessingJ. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene1982; 19:259–268
VosmanB., HellingwerfK.J. Molecular cloning and functional characterization of a recA analog from Pseudomonas stutzeri and construction of a P stutperi recA mutant. Antonie Eeeuwenhoek1991; 59:115–123
VosmanB., RauchP.J.G., WesterhoffH.V., HellingwerfK.J. Regulation of the expression of the Pseudomonas stutzeri rec A gene. Antonie Eeeuwenhoek1993; 63:55–62
WardhanH., McPhersonM.J., HarrisC.A., SharmaE., SastryG.R.K. Molecular analysis of the rec A gene of Agrobacterium tumejaciens C58. Gene1992; 121:133–136
ZultyJ.J., BarcakG.J. Structural organization, nucleotide sequence, and regulation of the Haemophilus influenzae rec- U gene. J Bacteriol1993; 175:7269–7281
The expression of the Acinetobacter calcoaceticus recA gene increases in response to DNA damage independently of RecA and of development of competence for natural transformation