Bishai W R, Smith H O, Barcak G J
Department of Medicine, Johns Hopkins Hospital, Baltimore, Maryland.
J Bacteriol. 1994 May;176(10):2914-21. doi: 10.1128/jb.176.10.2914-2921.1994.
Bacterial catalases are induced by exposure to peroxide (e.g., Escherichia coli katG) or entry into stationary phase (e.g., E. coli katE). To study regulatory systems in Haemophilus influenzae, we complemented an E. coli rpoS mutant, which is unable to induce katE in stationary phase, with a plasmid library of H. influenzae Rd- chromosomal DNA. Nineteen complementing clones with a catalase-positive phenotype were obtained and characterized after screening about 10(5) transformants. All carried the same structural gene for an H. influenzae catalase. The DNA sequence of this gene, called hktE, encodes a 508-amino-acid polypeptide with strong homology to eukaryotic catalases and E. coli katE. However, hktE is regulated like E. coli katG, with catalase activity increasing 10-fold and hktE mRNA levels increasing 4-fold upon exposure to ascorbic acid, which serves to generate hydrogen peroxide. Mutations in the known global regulatory genes of H. influenzae--crp, cya, and sxy--do not affect the inducibility of hktE. The hktE gene maps to a 225-kb segment of the H. influenzae chromosome in a region encoding resistance to spectinomycin.
细菌过氧化氢酶可通过暴露于过氧化物(如大肠杆菌katG)或进入稳定期(如大肠杆菌katE)诱导产生。为了研究流感嗜血杆菌中的调控系统,我们用流感嗜血杆菌Rd染色体DNA的质粒文库对处于稳定期时无法诱导katE的大肠杆菌rpoS突变体进行了互补。在筛选了约10⁵个转化体后,获得了19个具有过氧化氢酶阳性表型的互补克隆并进行了表征。所有克隆都携带相同的流感嗜血杆菌过氧化氢酶结构基因。这个名为hktE的基因的DNA序列编码一个508个氨基酸的多肽,与真核生物过氧化氢酶和大肠杆菌katE具有高度同源性。然而,hktE的调控方式与大肠杆菌katG类似,在暴露于用于产生过氧化氢的抗坏血酸后,过氧化氢酶活性增加10倍,hktE mRNA水平增加4倍。流感嗜血杆菌已知的全局调控基因crp、cya和sxy中的突变不影响hktE的诱导性。hktE基因定位于流感嗜血杆菌染色体的一个225 kb片段上,该区域编码对壮观霉素的抗性。