Azasugar-Containing Phosphorothioate Oligonucleotide (AZPSON) DBM-2198 Inhibits Human Immunodeficiency Virus Type 1 (HIV-1) Replication by Blocking HIV-1 gp120 without Affecting the V3 Region

Lee, Jinjoo; Byeon, Se Eun; Jung, Ju Yeol; Kang, Myeong-Ho; Park, Yu-Jin; Jung, Kyuwhan; Bae, Yong-Soo

doi:10.14348/molcells.2015.2129

Cited by 2 publications

(2 citation statements)

References 34 publications

(31 reference statements)

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“…UV absorption was monitored at 254. 4 . Ionization was set in positive mode, gas flow was 10 liters min Ϫ1 , nebulizer pressure was 30 lb/in 2 , drying gas temperature was 325°C, and capillary voltage was 3,100 V. Signals at m/z 597 (dGsA), 565 (dG H A), and 581 (dGA) were extracted to confirm the identities of the main peaks.…”

Section: Methodsmentioning

confidence: 99%

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Phosphorothioated DNA Is Shielded from Oxidative Damage

Mei

et al. 2019

Appl Environ Microbiol

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DNA is the carrier of genetic information. DNA modifications play a central role in essential physiological processes. Phosphorothioation (PT) modification involves the replacement of an oxygen atom on the DNA backbone with a sulfur atom. PT modification can cause genomic instability inSalmonella entericaunder hypochlorous acid stress. This modification restores hydrogen peroxide (H2O2) resistance in the catalase-deficientEscherichia coliHpx−strain. Here, we report biochemical characterization results for a purified PT modification protein complex (DndCDE) fromS. enterica. We observed multiplex oligomeric states of DndCDE by using native PAGE. This protein complex bound avidly to PT-modified DNA. DndCDE with an intact iron-sulfur cluster (DndCDE-FeS) possessed H2O2decomposition activity, with aVmaxof 10.58 ± 0.90 mM min−1and a half-saturation constant,K0.5S, of 31.03 mM. The Hill coefficient was 2.419 ± 0.59 for this activity. The protein’s activity toward H2O2was observed to be dependent on the intact DndCDE and on the formation of an iron-sulfur (Fe-S) cluster on the DndC subunit. In addition to cysteine residues that mediate the formation of this Fe-S cluster, other cysteine residues play a catalytic role. Finally, catalase activity was also detected in DndCDE fromPseudomonas fluorescensPf0-1. The data and conclusions presented suggest that DndCDE-FeS is a short-lived catalase. Our experiments also indicate that the complex binds to PT sites, shielding PT DNA from H2O2damage. This catalase shield might be able to extend from PT sites to the entire bacterial genome.IMPORTANCEDNA phosphorothioation has been reported in many bacteria. These PT-hosting bacteria live in very different environments, such as the human body, soil, or hot springs. The physiological function of DNA PT modification is still elusive. A remarkable property of PT modification is that purified genomic PT DNA is susceptible to oxidative cleavage. Among the oxidants, hypochlorous acid and H2O2are of physiological relevance for human pathogens since they are generated during the human inflammation response to bacterial infection. However, expression of PT genes in the catalase-deficientE. coliHpx−strain restores H2O2resistance. Here, we seek to solve this obvious paradox. We demonstrate that DndCDE-FeS is a short-lived catalase that binds tightly to PT DNA. It is thus possible that by docking to PT sites the catalase activity protects the bacterial genome against H2O2damage.

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Section: Methodsmentioning

confidence: 99%

“…PT DNA can more easily resist nuclease degradation (1). As a result, synthetic PT DNA has a broad range of uses in antisense technology (2)(3)(4).…”

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confidence: 99%

Phosphorothioated DNA Is Shielded from Oxidative Damage

Mei

et al. 2019

Appl Environ Microbiol

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Characterization of the PLP-dependent transaminase initiating azasugar biosynthesis

Arciola

Horenstein

2018

Biochemical Journal

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Biosynthesis of the azasugar 1-deoxynojirimycin (DNJ) critically involves a transamination in the first committed step. Here, we identify the azasugar biosynthetic cluster signature in SC2 (), homologous to that reported in FZB42 (), and report the characterization of the aminotransferase GabT1 (named from ). GabT1 from exhibits a specific activity of 4.9 nmol/min/mg at 30°C (pH 7.5), a somewhat promiscuous amino donor selectivity, and curvilinear steady-state kinetics that do not reflect the predicted ping-pong behavior typical of aminotransferases. Analysis of the first half reaction with l-glutamate in the absence of the acceptor fructose 6-phosphate revealed that it was capable of catalyzing multiple turnovers of glutamate. Kinetic modeling of steady-state initial velocity data was consistent with a novel hybrid branching kinetic mechanism which included dissociation of PMP after the first half reaction to generate the apoenzyme which could bind PLP for another catalytic deamination event. Based on comparative sequence analyses, we identified an uncommon His-Val dyad in the PLP-binding pocket which we hypothesized was responsible for the unusual kinetics. Restoration of the conserved PLP-binding site motif via the mutant H119F restored classic ping-pong kinetic behavior.

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Azasugar-Containing Phosphorothioate Oligonucleotide (AZPSON) DBM-2198 Inhibits Human Immunodeficiency Virus Type 1 (HIV-1) Replication by Blocking HIV-1 gp120 without Affecting the V3 Region

Cited by 2 publications

References 34 publications

Phosphorothioated DNA Is Shielded from Oxidative Damage

Phosphorothioated DNA Is Shielded from Oxidative Damage

Characterization of the PLP-dependent transaminase initiating azasugar biosynthesis

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