Toptchieva Anna, Sisson Gary, Bryden Louis J, Taylor Diane E, Hoffman Paul S
Department of Microbiology and Immunology, Dalhousie University, Sir Charles Tupper Medical Building, College Street, Halifax, Nova Scotia, Canada B3H 4H7.
Department of Medical Microbiology and Immunology, 1-28 Medical Science Building, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Microbiology (Reading). 2003 May;149(Pt 5):1285-1295. doi: 10.1099/mic.0.25981-0.
Tellurite resistance (Te(r)) is widespread in nature and it is shown here that the natural resistance of Proteus mirabilis to tellurite is due to a chromosomally located orthologue of plasmid-borne ter genes found in enteric bacteria. The P. mirabilis ter locus (terZABCDE) was identified in a screen of Tn5lacZ-generated mutants of which one contained an insertion in terC. The P. mirabilis terC mutant displayed increased susceptibility to tellurite (Te(s)) and complementation with terC carried on a multicopy plasmid restored high-level Te(r). Primer extension analysis revealed a single transcriptional start site upstream of terZ, but only with RNA harvested from bacteria grown in the presence of tellurite. Northern blotting and reverse transcriptase-PCR (RT-PCR) analyses confirmed that the ter operon was inducible by tellurite and to a lesser extent by oxidative stress inducers such as hydrogen peroxide and methyl viologen (paraquat). Direct and inverted repeat sequences were identified in the ter promoter region as well as motifs upstream of the -35 hexamer that resembled OxyR-binding sequences. Finally, the 390 bp intergenic promoter region located between orf3 and terZ showed no DNA sequence identity with any other published ter sequences, whereas terZABCDE genes exhibited 73-85 % DNA sequence identity. The ter operon was present in all clinical isolates of P. mirabilis and Proteus vulgaris tested and is inferred for Morganella and Providencia spp. based on screening for high level Te(r) and preliminary PCR analysis. Thus, a chromosomally located inducible tellurite resistance operon appears to be a common feature of the genus Proteus.
亚碲酸盐抗性(Te(r))在自然界广泛存在,本文表明奇异变形杆菌对亚碲酸盐的天然抗性是由于其染色体上存在与肠道细菌中质粒携带的ter基因同源的基因。在对Tn5lacZ产生的突变体进行筛选时鉴定出了奇异变形杆菌的ter基因座(terZABCDE),其中一个突变体在terC中存在插入。奇异变形杆菌terC突变体对亚碲酸盐的敏感性增加(Te(s)),用多拷贝质粒携带的terC进行互补可恢复高水平的Te(r)。引物延伸分析揭示在terZ上游有一个单一的转录起始位点,但仅在从在亚碲酸盐存在下生长的细菌中收获的RNA中存在。Northern印迹和逆转录酶PCR(RT-PCR)分析证实ter操纵子可被亚碲酸盐诱导,在较小程度上也可被氧化应激诱导剂如过氧化氢和甲基紫精(百草枯)诱导。在ter启动子区域鉴定出直接和反向重复序列以及类似于OxyR结合序列的-35六聚体上游的基序。最后,位于orf3和terZ之间的390 bp基因间启动子区域与任何其他已发表的ter序列没有DNA序列同一性,而terZABCDE基因表现出73-85%的DNA序列同一性。ter操纵子存在于所有测试的奇异变形杆菌和普通变形杆菌的临床分离株中,基于对高水平Te(r)的筛选和初步PCR分析推断摩根菌属和普罗威登斯菌属也存在。因此,染色体上定位的可诱导亚碲酸盐抗性操纵子似乎是变形杆菌属的一个共同特征。