Beaumont Hubertus J E, Lens Sylvia I, Reijnders Willem N M, Westerhoff Hans V, van Spanning Rob J M
BioCentrum Amsterdam, Department of Molecular Cell Physiology, Vrije Universiteit, de Boelelaan 1087, NL-1081 HV Amsterdam, The Netherlands.
Mol Microbiol. 2004 Oct;54(1):148-58. doi: 10.1111/j.1365-2958.2004.04248.x.
Production of nitric oxide (NO) and nitrous oxide (N(2)O) by ammonia (NH(3))-oxidizing bacteria in natural and man-made habitats is thought to contribute to the undesirable emission of NO and N(2)O into the earth's atmosphere. The NH(3)-oxidizing bacterium Nitrosomonas europaea expresses nitrite reductase (NirK), an enzyme that has so far been studied predominantly in heterotrophic denitrifying bacteria where it is involved in the production of these nitrogenous gases. The finding of nirK homologues in other NH(3)-oxidizing bacteria suggests that NirK is widespread among this group; however, its role in these nitrifying bacteria remains unresolved. We identified a gene, nsrR, which encodes a novel nitrite (NO(2) (-))-sensitive transcription repressor that plays a pivotal role in the regulation of NirK expression in N. europaea. NsrR is a member of the Rrf2 family of putative transcription regulators. NirK was expressed aerobically in response to increasing concentrations of NO(2) (-) and decreasing pH. Disruption of nsrR resulted in the constitutive expression of NirK. NsrR repressed transcription from the nirK gene cluster promoter (P(nir)), the activity of which correlated with NirK expression. Reconstruction of the NsrR-P(nir) system in Escherichia coli revealed that repression by NsrR was reversed by NO(2) (-) in a pH-dependent manner. The findings are consistent with the hypothesis that N. europaea expresses NirK as a defence against the toxic NO(2) (-) that is produced during nitrification.
在自然和人工环境中,氨氧化细菌产生一氧化氮(NO)和一氧化二氮(N₂O)被认为是导致这些气体向地球大气中不良排放的原因。氨氧化细菌欧洲亚硝化单胞菌表达亚硝酸还原酶(NirK),迄今为止,这种酶主要在异养反硝化细菌中进行研究,在这些细菌中它参与这些含氮气体的产生。在其他氨氧化细菌中发现nirK同源物表明NirK在这一类细菌中广泛存在;然而,它在这些硝化细菌中的作用仍未明确。我们鉴定出一个基因nsrR,它编码一种新型的对亚硝酸盐(NO₂⁻)敏感的转录阻遏物,该阻遏物在欧洲亚硝化单胞菌中对NirK表达的调控中起关键作用。NsrR是假定转录调节因子Rrf2家族的成员。随着NO₂⁻浓度增加和pH值降低,NirK在有氧条件下表达。nsrR的破坏导致NirK的组成型表达。NsrR抑制nirK基因簇启动子(Pnir)的转录,其活性与NirK表达相关。在大肠杆菌中重建NsrR - Pnir系统表明,NsrR的阻遏作用在pH依赖的方式下被NO₂⁻逆转。这些发现与欧洲亚硝化单胞菌表达NirK作为对硝化过程中产生的有毒NO₂⁻的防御这一假设一致。