An Insertion Sequence Prepares Pseudomonas putida S12 for Severe Solvent Stress
| Autor: | Jan Wery, Jasper Kieboom, Jan A.M. de Bont, Budi Hidayat |
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| Rok vydání: | 2001 |
| Předmět: |
Genotype
Molecular Sequence Data Population Adaptation Biological Gene Dosage Biochemistry Microbiology chemistry.chemical_compound Gene cluster Amino Acid Sequence Insertion sequence education Molecular Biology Gene education.field_of_study Base Sequence biology Pseudomonas putida Genetic Complementation Test Nucleic acid sequence Genetic Variation Biological Transport Cell Biology biology.organism_classification Molecular biology Toluene carbohydrates (lipids) Mutagenesis Insertional Phenotype chemistry DNA Transposable Elements ATP-Binding Cassette Transporters Bacteria |
| Zdroj: | Journal of Biological Chemistry. 276:5700-5706 |
| ISSN: | 0021-9258 |
| Popis: | The novel insertion sequence ISS12plays a key role in the tolerance of Pseudomonas putida S12 to sudden toluene stress. Under normal culturing conditions theP. putida S12 genome contained seven copies of ISS12. However, a P. putida S12 population growing to high cell density after sudden addition of a separate phase of toluene carried eight copies. The survival frequency of cells in this variant P. putida S12 population was 1000 times higher than in “normal” P. putida S12 populations. Analysis of the nucleotide sequence flanking the extra ISS12 insertion revealed integration into the srpS gene. srpSforms a gene cluster with srpR and both are putative regulators of the solvent resistance pump SrpABC. SrpABC makes a major contribution to solvent tolerance in P. putida S12 and is induced by toluene. The basal level of srp promoter activity in the P. putida S12 variant was seven times higher than in wild-type P. putida S12. Introduction of the intact srpRS gene cluster in the variant resulted in a dramatic decrease of survival frequency after a toluene shock. These findings strongly suggest that interruption of srpS by ISS12 up-regulates expression of the solvent pump, enabling the bacterium to tolerate sudden exposure to lethal concentrations of toxic solvents. We propose that P. putida S12 employs ISS12 as a mutator element to generate diverse mutations to swiftly adapt when confronted with severe adverse conditions. |
| Databáze: | OpenAIRE |
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