Wu J, Rosen B P
Department of Biochemistry, Wayne State University, School of Medicine, Detroit, Michigan 48201.
Mol Microbiol. 1993 May;8(3):615-23. doi: 10.1111/j.1365-2958.1993.tb01605.x.
The plasmid-encoded arsenical resistance (ars) operon produces resistance to trivalent and pentavalent salts of arsenic and antimony. The first gene in the operon, arsR, was previously shown to encode a repressor protein. A newly identified gene, arsD, is shown here to encode a regulatory protein, the ArsD protein. The gene was identified by construction of an in-frame fusion between the C-terminally truncated arsD gene and the coding region for the mature form of beta-lactamase (blaM). The native arsD gene product was overexpressed and radioactively labelled as a 13 kDa polypeptide. A frameshift mutation within the arsD gene resulted in elevated levels of expression of downstream ars genes. Co-expression of a wild-type arsD gene in trans with the operon containing the mutated arsD gene reduced expression of the downstream genes to wild-type levels. The presence of the arsD gene had no effect on the basal level of operon expression set by the arsR gene product, and the repression produced by the arsD gene product was not affected by inducers of the operon. The results indicate that the ArsD protein is an inducer-independent trans-acting regulatory protein.
质粒编码的抗砷(ars)操纵子可产生对砷和锑的三价及五价盐的抗性。操纵子中的第一个基因arsR,先前已证明其编码一种阻遏蛋白。本文显示,一个新鉴定的基因arsD编码一种调节蛋白,即ArsD蛋白。该基因是通过构建C端截短的arsD基因与成熟形式的β-内酰胺酶(blaM)编码区之间的读码框融合体而鉴定出来的。天然arsD基因产物以13 kDa多肽的形式过表达并进行放射性标记。arsD基因内的一个移码突变导致下游ars基因的表达水平升高。野生型arsD基因与含有突变arsD基因的操纵子共表达,可使下游基因的表达降至野生型水平。arsD基因的存在对arsR基因产物设定的操纵子基础表达水平没有影响,并且arsD基因产物产生的阻遏作用不受操纵子诱导剂的影响。结果表明,ArsD蛋白是一种不依赖诱导剂的反式作用调节蛋白。