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The merR regulatory gene in Thiobacillus ferrooxidans is spaced apart from the mer structural genes
Author(s) -
Inoue C.,
Sugawara K.,
Kusano T.
Publication year - 1991
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1991.tb01979.x
Subject(s) - orfs , biology , gene , genetics , open reading frame , homology (biology) , plasmid , nucleic acid sequence , transposable element , structural gene , peptide sequence , escherichia coli , microbiology and biotechnology , genome
Summary Two distinct merR genes, which regulate expression of the mercuric ion resistance gene ( mer ), of Thiobacillus ferrooxidans strain E‐15 have been cloned, sequenced and termed merR 1 , and merR 2 . As a result of gene walking around two merR genes, it was found that these two genes were quite close in distance. The nucleotide sequence of the region (5001 base pairs; PstI‐EcoRI fragment ) containing the merR genes was determined. Between the two merR genes, there were five potential open reading frames (ORFs). Two of these were identified as merC genes, and the other three as ORFs 1 to 3. ORFs 1 to 3 show significant homology to merA, tnsA from transposon Tn7, and merA , respectively. Both merR genes consist of a 408 bp ORF coding for 135 amino acids. Their gene products, MerRl and MerR2, differed at three amino acid positions, and shared 56‐57% and 32‐38% identity with the MerRs from other Gram‐negative and Gram‐positive bacteria, respectively. Competitive primer extension analysis revealed that both regulatory genes were expressed in the host cells. These merR genes were located more than 6 kb from either end of the mer structural genes (merC‐merA ). This is the first example of merR being separated from the mer structural genes. The two merC genes, each of which coded for a 140‐amino‐acid protein, appeared to be functionally active because Escherichia coli cells carrying these merC geneson plasmid vectors showed hypersensitivity to HgCl 2 . However, ORFs 1 and 3, which were homologous to merA , seemed to be inactive both structurally and enzymatically. The gene arrangement in this region took on a mirror image, with the truncated tnsA as the symmetrical centre. It is suggested that the Tn7‐like factor may have participated in gene duplication events of the mer region, and in its chromosomal integration.