Summary
The recA gene of Pseudomonas aeruginosa has been isolated and its nucleotide sequence has been determined. The coding region of the recA gene has 1038 bp specifying 346 amino acids. The recA protein of P. aeruginosa showed a striking homology with that of Escherichia coli except for the carboxy-terminal region both at the nucleotide and amino acid level. The recA +-carrying plasmids restored the UV sensitivity and recombination ability of several rec mutants of P. aeruginosa. The precise location of the recA gene on the chromosome was deduced from the analysis of R′ plasmids.
Similar content being viewed by others
References
Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–452
Bagdasarian M, Lurz R, Rückert B, Franklin FCH, Bagdasarian MM, Frey J, Timmis KN (1981) Specific-purpose plasmid cloning vectors II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene 16:237–247
Bagdasarian MM, Amann E, Lurz R, Rückert B, Bagdasarian M (1983) Activity of the hybrid trp-lac(tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad host range, controlled-expression vectors. Gene 26:273–282
Better M, Helinski DR (1983) Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol 155:311–316
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC (1984) A rapid, sensitive method for detection of alkaline phosphataseconjugated anti-antibody on Western blots. Anal Biochem 136:175–179
Bolivar F, Rodriguez RL, Greene PJ, Betlach MJ, Heynecker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
Boyer HW, Roulland-Dussoix D (1969) A Complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Burgess RR, Jendrisak JJ (1975) A procedure for the rapid, largescale purification of Escherichia coli DNA-dependent RNA polymerase involving polymin P precipitation and DNA-cellulose chromatography. Biochemistry 14:4634–4638
Chandler PM, Krishnapillai V (1974) Isolation and properties of recombination-deficient mutants of Pseudomonas aeruginosa. Mutat Res 23:15–23
Clark AJ, Margulies AD (1965) Isolation and characterization of recombination deficient mutants of Escherichia coli. Proc Natl Acad Sci USA 53:451–459
Eitner G, Adler B, Lanzov VA, Hofemeister J (1982) Interspecies recA protein substitution in Escherichia coli and Proteus mirabilis. Mol Gen Genet 185:481–486
Früh R, Watson JM, Haas D (1983) Construction of recombination-deficient strains of Pseudomonas aeruginosa. Mol Gen Genet 191:334–337
Gill DR, Hatfull GF, Salmond GPC (1986) A new cell division operon in Escherichia coli. Mol Gen Genet 205:134–145
Goldberg I, Mekalanos JJ (1986) Cloning of the Vibrio cholerae recA gene and construction of a Vibrio cholerae recA mutant. J Bacteriol 165:715–722
Haas D, Holloway BW (1976) R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet 144:243–251
Hamood AN, Pettis GS, Parker CD, McIntosh MA (1986) Isolation and characterization of the Vibrio cholerae recA gene. J Bacteriol 167:375–378
Harayama S, Oguchi T, Iino T (1984) Does Tn10 transpose via the cointegrate molecule? Mol Gen Genet 194:444–450
Hattori M, Sakaki Y (1986) Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 152:232–238
Higgins CF, Hiles ID, Whalley K, Jamieson DJ (1985) Nucleotide binding bymembrane components of bacterial periplasmic binding protein-dependent transport system. EMBO J 4:1033–1040
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Horii T, Ogawa T, Ogawa H (1980) Organization of the recA gene of Escherichia coli. Proc Natl Acad Sci USA 77:313–317
Kawashima H, Horii T, Ogawa T, Ogawa H (1984) Functional domains of Escherichia coli recA protein deduced from the mutational sites in the gene. Mol Gen Genet 193:288–292
Keener SL, McNamee KP, McEntee K (1984) Cloning and characterization of recA genes from Proteus vulgaris, Erwinia carotovora, Shigella flexneri and Escherichia coli B/r. J Bacteriol 160:153–160
Kogoma T, Skarstad K, Boye E, Meyenburg K von, Steen HB (1985) RecA protein acts at the initiation of stable DNA replication in rnh mutants of Escherichia coli K12. J Bacteriol 163:439–444
Kokjohn TA, Miller RV (1985) Molecular cloning and characterization of the recA gene of Pseudomonas aeruginosa PAO. J Bacteriol 163:568–572
Kuramitsu S, Hamaguchi K, Tachibana H, Horii T, Ogawa T, Ogawa H (1984) Cysteinyl residues of Escherichia coli recA protein. Biochemistry 23:2363–2367
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning-a laboratory mannual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Masaki H, Ohta T (1985) Colicin E3 and its immunity genes. J Mol Biol 182:217–227
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with basespecific chemical cleavages. Methods Enzymol 65:499–560
Miyata T, Miyazawa S, Yasunaga T (1979) Two types of amino acid substitutions in protein evolution. J Mol Evol 12:219–236
Ogawa H, Ogawa T (1986) General recombination: Functions and structure of recA protein. Adv Biophys 21:135–148
Ogawa T, Wabiko H, Tsurimoto T, Horii T, Masukata H, Ogawa H (1979) Characteristics of purified recA protein and the regulation of its synthesis in vivo. Cold Spring Harbor Symp Quant Biol 43:909–915
Ohman DE, West MA, Flynn JL, Goldberg JB (1985) Method for gene replacement in Pseudomonas aeruginosa used in construction of recA mutants: recA-independent instability of alginate production. J Bacteriol 162:1068–1074
Rüther U, Koenen M, Otto K, Muller-Hill B (1981) pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucleic Acids Res 9:4087–4098
Sancar A, Stachelek C, Konigsberg W, Rupp WD (1980) Sequences of the recA gene and protein. Proc Natl Acad Sci USA 77:2611–2615
Sano Y, Kageyama M (1977) Transformation of Pseudomonas aeruginosa by plasmid DNA. J Gen Appl Microbiol 23:183–186
Sano Y, Kageyama M (1981) Purification and properties of an S-type pyocin, pyocin AP41. J Bacteriol 146:733–739
Sano Y, Kageyama M (1984) Genetic determinant of pyocin AP41 as an insert in the Pseudomonas aeruginosa chromosome. J Bacteriol 158:562–570
Shinomiya T, Shiga S, Kageyama M (1983) Genetic determinant of pyocin R2 in Pseudomonas aeruginosa PAO I. Localization of the pyocin R2 gene cluster between the trpCD and trpE genes. Mol Gen Genet 189:375–381
Smith DE, Fisher PA (1984) Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: Application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots. J Cell Biol 99:20–28
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the α and β subunits of ATP synthase, myocin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951
Walker GC (1984) Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev 48:60–93
Wang W-B, Tessman ES (1985) Evidence that the recA441 (tif-1) mutant of Escherichia coli K-12 contains a thermosensitive intragenic suppressor of recA constitutive protease activity. J Bacteriol 163:407–409
Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev 40:869–907
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Author information
Authors and Affiliations
Additional information
Communicated by M. Takanami
Rights and permissions
About this article
Cite this article
Sano, Y., Kageyama, M. The sequence and function of the recA gene and its protein in Pseudomonas aeruginosa PAO. Mol Gen Genet 208, 412–419 (1987). https://doi.org/10.1007/BF00328132
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00328132