Nitrosative damage to free and zinc‐bound cysteine thiols underlies nitric oxide toxicity in wild‐type <i>Borrelia burgdorferi</i>

Bourret, Travis J.; Boylan, Julie A.; Lawrence, Kevin A.; Gherardini, Frank C.

doi:10.1111/j.1365-2958.2011.07691.x

Cited by 43 publications

(67 citation statements)

References 51 publications

(84 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several enzymes previously shown to be inhibited by NO could be identified by the buildup of their substrates or upstream intermediates. For instance, fructose-1,6-bisphosphate aldolase is a zinc-dependent glycolytic enzyme demonstrated to be NO sensitive in Borrelia burgdorferi (14). In our study, upstream glycolytic intermediates, such as glucose, glucose-6-phosphate, and fructose-1,6-bisphosphate, all accumulated in the presence of NO, whereas downstream metabolites, such as 2-phosphoglycerate and 3-phosphoglycerate, decreased (see Fig.…”

Section: Nnrs Is Important For Resistance To No But Does Not Remove Nosupporting

confidence: 54%

“…DNICs have also been shown to mediate the formation of nitrosothiols, another form of nitrosative stress that inhibits thiolcontaining proteins (10). Through this mechanism and others, NO has been shown to inhibit a few enzymes in vitro, including such central metabolic enzymes as aconitase (11), dihydroxyacid dehydratase (12), alpha-ketoglutarate dehydrogenase (13), fructose-1,6-bisphosphate aldolase (14), argininosuccinate synthase (15), and components of the respiratory chain (16,17). However, these enzymes have largely been studied in isolation, and there has not been any comprehensive study on the effects of NO on bacterial metabolism.…”

mentioning

confidence: 99%

See 1 more Smart Citation

A Novel Protein Protects Bacterial Iron-Dependent Metabolism from Nitric Oxide

et al. 2013

View full text Add to dashboard Cite

Reactive nitrogen species (RNS), in particular nitric oxide (NO), are toxic to bacteria, and bacteria have mechanisms to allow growth despite this stress. Understanding how bacteria interact with NO is essential to understanding bacterial physiology in many habitats, including pathogenesis; however, many targets of NO and enzymes involved in NO resistance remain uncharacterized. We performed for the first time a metabolomic screen on NO-treated and -untreated bacteria to define broadly the effects of NO on bacterial physiology, as well as to identify the function of NnrS, a previously uncharacterized enzyme involved in defense against NO. We found many known and novel targets of NO. We also found that iron-sulfur cluster enzymes were preferentially inhibited in a strain lacking NnrS due to the formation of iron-NO complexes. We then demonstrated that NnrS is particularly important for resistance to nitrosative stress under anaerobic conditions. Our data thus reveal the breadth of the toxic effects of NO on metabolism and identify the function of an important enzyme in alleviating this stress. Vibrio cholerae causes cholera, a severe watery diarrhea responsible for millions of cases and thousands of deaths each year (Centers for Disease Control and Prevention [http://www.cdc .gov]). It is not, however, a member of the Enterobacteriaceae-its natural habitat is aquatic. It is thought that during most of its life cycle, when not infecting humans, V. cholerae resides in association with zooplankton, forming biofilms on the chitinous surfaces of crustaceans (1, 2). Thus, V. cholerae must display metabolic flexibility in order to thrive in these two different environments and respond to the different metabolic challenges therein.A commonly encountered metabolic stress for pathogenic and nonpathogenic bacteria is the presence of reactive nitrogen species (RNS), in particular the well-studied molecule nitric oxide (NO). NO is formed as a by-product of nitrogen metabolism for many bacteria as an intermediate in denitrification (3), as well as from dedicated nitric oxide synthases (NOSs) in both bacteria and eukaryotes (4, 5), and is present in micromolar concentrations in some bacterial biofilms (6). NO can also be formed by chemical decomposition of nitrite in acid environments, such as the human stomach (7). NO is also a prominent component of the mammalian innate immune system, part of a battery of reactive oxygen species (ROS) and RNS produced by phagocytes when they encounter bacteria (7).The mechanisms whereby NO inhibits bacterial growth are diverse, but one of the most important properties of NO is its ability to bind iron and form dinitrosyl iron complexes (DNICs) bound to iron-sulfur cluster proteins, inhibiting their function (8, 9). DNICs have also been shown to mediate the formation of nitrosothiols, another form of nitrosative stress that inhibits thiolcontaining proteins (10). Through this mechanism and others, NO has been shown to inhibit a few enzymes in vitro, including such central metabolic enzymes as ...

show abstract

Section: Nnrs Is Important For Resistance To No But Does Not Remove Nosupporting

confidence: 54%

mentioning

confidence: 99%

A Novel Protein Protects Bacterial Iron-Dependent Metabolism from Nitric Oxide

et al. 2013

View full text Add to dashboard Cite

show abstract

“…A previous study has reported that disruption of B. burgdorferi uvrB and uvrC results in decreased survival following 4 h exposure to the NO donor DEA/NO (43). However, in another study, using a different strain of B. burgdorferi exposed for 1 h to different sources of NO, disruption of uvrA did not result in significantly increased sensitivity (38).…”

Section: Figmentioning

confidence: 82%

“…The nucleotide excision repair pathway is required for survival to reactive nitrogen stress, which is known to affect the infectivity of pathogens (54). Although previous studies have described the role of some NER genes for resistance to DNA damage in B. burgdorferi (38,43), our current study is the first time that the importance of the NER pathway for infectivity has been tested. In order to detect a difference in infectivity between each mutant, mice were infected with 10 3 spirochetes at each of two sites at a dose close to the minimum infectious dose required for infection of all mice and for dissemination to all target organs tested with strain B31 (55).…”

Section: Figmentioning

confidence: 93%

“…Sensitivity to reactive nitrogen species (RNS) damage was determined as previously described (43). Cultures were grown to a density of 1 ϫ 10 7 to 5 ϫ 10 7 cells ml Ϫ1 , and then spirochetes were pelleted and resuspended in culture medium at 5 ϫ 10 7 spirochetes per ml with 2.5 mM diethylamine (DEA) NONOate diethylammonium salt (DEA/NO; Sigma-Aldrich, Oakville, ON, Canada) or with culture medium only.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The Nucleotide Excision Repair System of Borrelia burgdorferi Is the Sole Pathway Involved in Repair of DNA Damage by UV Light

Hardy

Chaconas

2013

J Bacteriol

View full text Add to dashboard Cite

To survive and avoid accumulation of mutations caused by DNA damage, the genomes of prokaryotes encode a variety of DNA repair pathways most well characterized in Escherichia coli. Some of these are required for the infectivity of various pathogens. In this study, the importance of 25 DNA repair/recombination genes for Borrelia burgdorferi survival to UV-induced DNA damage was assessed. In contrast to E. coli, where 15 of these genes have an effect on survival of UV irradiation, disruption of recombinational repair, transcription-coupled repair, methyl-directed mismatch correction, and repair of arrested replication fork pathways did not decrease survival of B. burgdorferi exposed to UV light. However, the disruption of the B. burgdorferi nucleotide excision repair (NER) pathway (uvrA, uvrB, uvrC, and uvrD) resulted in a 10-to 1,000-fold increase in sensitivity to UV light. A functional NER pathway was also shown to be required for B. burgdorferi resistance to nitrosative damage. Finally, disruption of uvrA, uvrC, and uvrD had only a minor effect upon murine infection by increasing the time required for dissemination.

show abstract

Physiological Roles of Nitrite and Nitric Oxide in Bacteria: Similar Consequences from Distinct Cell Targets, Protection, and Sensing Systems

Guo

Gao

2021

Advanced Biology

View full text Add to dashboard Cite

sodium nitrite) is currently in a phase 2b clinical trial as a drug for pulmonary hypertension treatment on the basis of the finding that it could be safely applied to reach millimolar concentrations in the airway surface liquid. [6] In the broad nitrogen biogeochemical cycle, nitrite serves as the central molecule linking nitrate (NO 3 − ) to dinitrogen gas (N 2 ) or ammonia (NH 3 ). [7] As a nitrogen oxo-anion, nitrite per se is crucial to life on earth because it participates in diverse key biological processes, especially in bacteria because they are renowned for their ability to carry out biological transformation of nitrite (Maia and Moura, 2014). Nitrite can be oxidized to nitrate or reduced to various nitrogen species such as nitric oxide (NO), nitrous oxide (N 2 O), N 2 , and NH 3 . [8] Although nitrite transformation is regarded as an effective means for detoxification, it brings out new threats. [7] Apart from an intermediate in the nitrogen cycle, NO, whose physiological concentrations are suggested to be up to ≈5 nM in cells, is a well-recognized bacteriostatic agent the same as nitrite. [9] It should be noted that concentrations of NO generated from nitrite reduction vary drastically in different bacteria, for example, less than 20 nM and up to 38 µM in cultures of two denitrifiers Paracoccus denitrificans and Agrobacterium tumefaciens, respectively. [10] In bacteria, a variety of enzymes are implicated in generation of NO from nitrite, and consistently, multiple enzymes contribute to transformation of NO to other nitrogen species for detoxification. [7,11] It is worth noting that in contrast to nitrite, whose transformation is exclusively conducted by enzymes involved in the nitrogen biogeochemical cycle, NO can be generated and scavenged by proteins outside the cycle, bacterial NO synthases (bNOSs) and flavohemoglobin Hmp, respectively. [12] Both nitrite and NO are bacteriostatic agents due to their ability to inhibit protein activity. In vitro studies show that nitrite and NO display similar, albeit not identical, biochemical properties, and therefore proteins susceptible to them are similar, mainly those containing redox-active centers such as heme, iron-sulfur clusters, mono-/di-nuclear iron, thiol, and so on. [13] Consistently, most of cellular targets identified in vivo are metabolic and respiratory enzymes depending on their redox-active centers for catalysis and/or oxi-reduction. [11] Despite this, cellular targets of nitrite and NO in any given bacterial species are Nitrite and nitric oxide (NO) are two active nitrogen oxides that display similar biochemical properties, especially when interacting with redox-sensitive proteins (i.e., hemoproteins), an observation serving as the foundation of the notion that the antibacterial effect of nitrite is largely attributed to NO formation. However, a growing body of evidence suggests that they are largely treated as distinct molecules by bacterial cells. Although both nitrite and NO are formed and decomposed by enzymes participating in the trans...

show abstract

Nitrosative damage to free and zinc‐bound cysteine thiols underlies nitric oxide toxicity in wild‐type Borrelia burgdorferi

Cited by 43 publications

References 51 publications

A Novel Protein Protects Bacterial Iron-Dependent Metabolism from Nitric Oxide

A Novel Protein Protects Bacterial Iron-Dependent Metabolism from Nitric Oxide

The Nucleotide Excision Repair System of Borrelia burgdorferi Is the Sole Pathway Involved in Repair of DNA Damage by UV Light

Physiological Roles of Nitrite and Nitric Oxide in Bacteria: Similar Consequences from Distinct Cell Targets, Protection, and Sensing Systems

Contact Info

Product

Resources

About