Characterization of the Pseudomonas aeruginosa recA analog and its protein product: rec-102 is a mutant allele of the P. aeruginosa PAO recA gene
We cloned a 2.3-kilobase-pair fragment of the Pseudomonas aeruginosa PAO chromosome which is capable of complementing recA mutations of Escherichia coli. The recA-complementing activity was further localized to a 1.5-kilobase-pair PvuII-HindIII fragment. Southern blot analysis under conditions of high stringency indicated that DNA sequence homology is shared by the E. coli recA gene and the P. aeruginosa recA analog. The cloned recA analog was shown to restore resistance to methyl methanesulfonate, nitrofurantoin, and UV irradiation to E. coli recA mutants. Upon introduction of the cloned P. aeruginosa gene, these mutants regained recombination proficiency in HfrH-mediated conjugation and the ability to induce lambda prophages and SOS functions (din gene transcription) after exposure to DNA-damaging agents. Lambda prophage carrying a cI ind mutation was not inducible, suggesting that the mechanism of induction of these SOS functions by the P. aeruginosa RecA analog is similar to that by the activated E. coli RecA protein. The product of the recA analog was identified in minicells as a protein of approximately 47,000 daltons. Western blot analysis using anti-E. coli RecA antibody demonstrated that this protein is antigenically cross-reactive with the E. coli recA protein. The recA-containing fragment was cloned into the broad-host-range vector pCP13 and introduced into Rec- strains of P. aeruginosa containing the rec-102 allele. The plasmid was shown to restore recombination proficiency in FP5-mediated conjugations and to restore resistance to UV irradiation and methyl methanesulfonate to these Rec- mutants. It was shown that a wild-type allele of rec-102 is necessary for UV-mediated induction of D3 and F116 prophages. The cloned recA analog restored the UV inducibility of these prophages in rec-102 mutants.
- Research Organization:
- Loyola Univ. of Chicago, Maywood, IL
- OSTI ID:
- 6791681
- Journal Information:
- J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 4; ISSN JOBAA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560130 -- Radiation Effects on Microorganisms
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANTIGEN-ANTIBODY REACTIONS
BACTERIA
BACTERIOPHAGES
CELL CONSTITUENTS
CLONING
DNA
DNA SEQUENCING
ELECTROMAGNETIC RADIATION
ESCHERICHIA COLI
ESTERS
GENE RECOMBINATION
GENES
METHYL METHANESULFONATE
MICROORGANISMS
MUTAGENS
MUTATIONS
NITRO COMPOUNDS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PARASITES
PLASMIDS
PROTEINS
PSEUDOMONAS
RADIATIONS
RADIOSENSITIVITY
RECOMBINANT DNA
STRUCTURAL CHEMICAL ANALYSIS
SULFONIC ACID ESTERS
TRANSCRIPTION
ULTRAVIOLET RADIATION
VIRUSES