Genetic analysis of epidermin biosynthetic genes and epidermin-negative mutants of Staphylococcus epidermidis
| Autor: | Karl-Dieter Entian, Uschi Schneider, Ralf Rosenstein, Friedrich Götz, Schnell Norbert, Johannes Augustin, Wieland Bernd, Germar Engelke |
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| Rok vydání: | 1992 |
| Předmět: |
DNA
Bacterial Molecular Sequence Data Mutant Biology medicine.disease_cause Peptides Cyclic Polymerase Chain Reaction Biochemistry Open Reading Frames Plasmid Bacteriocins Staphylococcus epidermidis medicine Amino Acid Sequence Gene Genetics Mutation Base Sequence Structural gene Nucleic acid sequence Promoter Gene Expression Regulation Bacterial Molecular biology Anti-Bacterial Agents Open reading frame Genes Bacterial Mutagenesis Peptides Plasmids |
| Zdroj: | European Journal of Biochemistry. 204:1149-1154 |
| ISSN: | 1432-1033 0014-2956 |
| DOI: | 10.1111/j.1432-1033.1992.tb16740.x |
| Popis: | Epidermin is produced by Staphylococcus epidermidis Tu3298 which harbors the 54-kb plasmid, pTu32. The plasmid contains not only the epidermin structural gene epiA, but also a flanking DNA region which is necessary for epidermin biosynthesis. The DNA sequence of this region revealed, in addition to epiA, five additional open reading frames, epiB, C, D, Q and P [Schnell, N., Engelke, G., Augustin J., Rosenstein, R., Ungermann, V., Gotz, F. & Entian, K.-D. (1992) Eur. J. Biochem. 204, 57-68]. We isolated a number of stable mutants from strain Tu3298 which are unable to produce biologically active epidermin. Complementation studies using the newly constructed staphylococcal plasmid vectors pT181mcs and pCU1 led to their classification as epiA, epiB, epiC or epiD mutants. Furthermore, evidence is presented that epiB lacks its own promoter and is co-transcribed from the epiA promoter. There is evidence that epiC and D possess their own promoters. Although epiQ and epiP mutants were not isolated, it could be shown by heterologous gene expression in S. carnosus and S. xylosus that the corresponding DNA region is involved in epidermin biosynthesis. We can not exclude the possibility that, in addition to the four open reading frames, epiA, B, C, D, and the DNA region comprising epiQ and P, host-encoded functions are necessary for epidermin production. Thus, the genetic information for epidermin biosynthesis in S. carnosus and S. xylosus is located on an 8-kb DNA fragment of pTu32. A further characterization of the two epiA mutants revealed that in both mutants, the preepidermin nucleotide sequence was changed. In one mutant, the mutation led to a substitution of Ser3 by Asn; in the other of Gly10 by Glu. |
| Databáze: | OpenAIRE |
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