Overton Tim W, Whitehead Rebekah, Li Ying, Snyder Lori A S, Saunders Nigel J, Smith Harry, Cole Jeff A
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
J Biol Chem. 2006 Nov 3;281(44):33115-26. doi: 10.1074/jbc.M607056200. Epub 2006 Sep 5.
Neisseria gonorrhoeae survives anaerobically by reducing nitrite to nitrous oxide catalyzed by the nitrite and nitric oxide reductases, AniA and NorB. P(aniA) is activated by FNR (regulator of fumarate and nitrate reduction), the two-component regulatory system NarQ-NarP, and induced by nitrite; P(norB) is induced by NO independently of FNR by an uncharacterized mechanism. We report the results of microarray analysis, bioinformatic analysis, and chromatin immunoprecipitation, which revealed that only five genes with readily identified NarP-binding sites are differentially expressed in narP(+) and narP strains. These include three genes implicated in the truncated gonococcal denitrification pathway: aniA, norB, and narQ. We also report that (i) nitrite induces aniA transcription in a narP mutant; (ii) nitrite induction involves indirect inactivation by nitric oxide of a gonococcal repressor, NsrR, identified from a multigenome bioinformatic study; (iii) in an nsrR mutant, aniA, norB, and dnrN (encoding a putative reactive nitrogen species response protein) were expressed constitutively in the absence of nitrite, suggesting that NsrR is the only NO-sensing transcription factor in N. gonorrhoeae; and (iv) NO rather than nitrite is the ligand to which NsrR responds. When expressed in Escherichia coli, gonococcal NarQ and chimaeras of E. coli and gonococcal NarQ are ligand-insensitive and constitutively active: a "locked-on" phenotype. We conclude that genes involved in the truncated denitrification pathway of N. gonorrhoeae are key components of the small NarQP regulon, that NarP indirectly regulates P(norB) by stimulating NO production by AniA, and that NsrR plays a critical role in enabling gonococci to evade NO generated as a host defense mechanism.
淋病奈瑟菌通过亚硝酸还原酶和一氧化氮还原酶(AniA和NorB)催化将亚硝酸盐还原为一氧化二氮来进行厌氧生存。P(aniA)由FNR(延胡索酸和硝酸盐还原调节因子)、双组分调节系统NarQ-NarP激活,并由亚硝酸盐诱导;P(norB)由NO通过一种未明确的机制独立于FNR诱导。我们报告了微阵列分析、生物信息学分析和染色质免疫沉淀的结果,这些结果表明,在narP(+)和narP菌株中,只有五个具有易于识别的NarP结合位点的基因表达存在差异。其中包括三个与截短的淋球菌反硝化途径相关的基因:aniA、norB和narQ。我们还报告了:(i)亚硝酸盐在narP突变体中诱导aniA转录;(ii)亚硝酸盐诱导涉及从多基因组生物信息学研究中鉴定出的淋球菌阻遏物NsrR被一氧化氮间接失活;(iii)在nsrR突变体中,aniA、norB和dnrN(编码一种假定的活性氮物种反应蛋白)在没有亚硝酸盐的情况下组成性表达,这表明NsrR是淋病奈瑟菌中唯一的NO感应转录因子;以及(iv)NsrR响应的配体是NO而不是亚硝酸盐。当在大肠杆菌中表达时,淋球菌NarQ以及大肠杆菌和淋球菌NarQ的嵌合体对配体不敏感且组成性激活:一种“锁定”表型。我们得出结论,参与淋病奈瑟菌截短反硝化途径的基因是小NarQP调节子的关键组成部分,NarP通过刺激AniA产生NO间接调节P(norB),并且NsrR在使淋球菌逃避作为宿主防御机制产生的NO方面发挥关键作用。