GENETIC-ANALYSIS OF EPIDERMIN BIOSYNTHETIC GENES AND EPIDERMIN-NEGATIVE MUTANTS OF STAPHYLOCOCCUS-EPIDERMIDIS
GENETIC-ANALYSIS OF EPIDERMIN BIOSYNTHETIC GENES AND EPIDERMIN-NEGATIVE MUTANTS OF STAPHYLOCOCCUS-EPIDERMIDIS
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DOI：
10.1111/j.1432-1033.1992.tb16740.x
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发表时间：
1992-03-15
期刊：
影响因子：
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通讯作者：
GOTZ, F
中科院分区：
文献类型：
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作者：
AUGUSTIN, J;ROSENSTEIN, R;GOTZ, F
表皮素由表皮葡萄球菌Tu 3298产生，其携带54-kb质粒pTu 32。该质粒不仅含有表皮素结构基因epiA，而且含有表皮素生物合成所必需的侧翼DNA区域。该区域的DNA序列显示，除了epiA之外，还有五个额外的开放阅读框，epiB、C、D、Q和P [Schnell，N.，Engelke，G.，奥古斯丁·J罗森斯坦河Ungermann，V.，Gotz，F. &恩田。K.- D.（1992）Eur. J. Biochem.204，57-68]。我们从菌株Tu 3298中分离出许多稳定的突变体，这些突变体不能产生生物活性的表皮素。使用新构建的葡萄球菌质粒载体pT 181 mcs和pCU 1的互补研究导致它们被分类为epiA、epiB、epic或epiD突变体。此外，有证据表明epiB缺乏自己的启动子，并从epiA启动子共转录。有证据表明epiC和D拥有自己的启动子。虽然没有分离到epiQ和epiP突变体，但可以通过异源基因在S. carnosus和S.木糖中相应的DNA区域参与表皮素的生物合成。我们不能排除这样的可能性，即除了四个开放阅读框epiA、B、C、D和包含epiQ和P的DNA区域之外，宿主编码的功能对于表皮蛋白的产生是必需的。因此，表皮素生物合成的遗传信息在S。carnosus和S.木糖位于pTu 32的8-kb DNA片段上。两个epiA突变体的进一步表征表明，在这两个突变体中，preepidermin核苷酸序列发生了变化。在一个突变体中，突变导致丝氨酸3被天冬酰胺取代;在另一个突变体中，Gly 10被Glu取代。
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.