Synthesis of riboguanosine pentaphosphate ppprGpp (Magic Spot II) via a phosphotriester approach

Schattenkerk, Cecil; Wreesmann, C. T. J.; Marel, G.A. van der; Boom, Jacques H. van

doi:10.1093/nar/13.10.3635

Cited by 17 publications

(11 citation statements)

References 11 publications

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“…Syntheses-The synthesis of nucleoside 3Ј-polyphosphates has received considerably less attention than have those of the corresponding 5Ј-polyphosphates because the former are more difficult to synthesize and because their biological roles are not as well described (23)(24)(25)(26)(27)(28)(29)(30)(31). To our knowledge the only reported chemical synthesis of 2Ј-deoxy-nucleoside 3Ј-triphosphates was that by Josse and Moffatt (28) and proceeded by homologation of the corresponding nucleoside 3Ј-monophosphate by use of phosphoromorpholidate.…”

Section: Resultsmentioning

confidence: 99%

“…The major drawback with this method is that the synthesis of the phosphoromorpholidates requires easy accessibility to the corresponding nucleoside 3Јmonophosphate. Van Boom and colleagues prepared nucleoside 3Ј-diphosphates in the thymidine and guanosine series by a phosphotriester approach that involved the use of an activated morpholinylphosphonate (27,30,31). Unfortunately, such a procedure cannot be applied when the base of the nucleotide is adenine, the 6-amino group being more reactive toward phosphorylating agents than is the 3Ј-hydroxyl of the nucleoside moiety (32,33).…”

Section: Resultsmentioning

confidence: 99%

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Inhibition of Adenylyl Cyclase by a Family of Newly Synthesized Adenine Nucleoside 3′-Polyphosphates

Désaubry

Shoshani

Johnson

1996

Journal of Biological Chemistry

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The synthesis of a number of adenine nucleoside 3'-polyphosphates has been devised via a phosphotriester approach that combines the method of alkoxide activation with the use of 2,2,2-tribromoethyl phosphoromorpholinochloridate as a phosphorylating agent. The family of compounds included 3'ADP, 3'ATP, 2'-deoxy-3'ADP, 2'-deoxy-3'ATP, 2',5'-dideoxy-3'ADP, and 2',5'-dideoxy-3'ATP. Potency as inhibitors of adenylyl cyclases followed the order: 3'-mono- < 3'-di- < 3'-triphosphate and adenosine (Ado) < 2'-d-Ado < 2',5'-dd-Ado derivatives, with 2',5'-dideoxy-3'ATP exhibiting an IC50 of approximately 40 nM. This order was maintained with purified and recombinant forms of the type I enzyme. The nucleoside 3'-phosphates caused noncompetitive inhibition of the type I adenylyl cyclase from bovine brain, consistent with inhibition via the P-site. Inhibition was not due to hydrolytic products because this was minimal and inhibition kinetics by inorganic polyphosphates were inconsistent with those caused by the nucleoside 3'-polyphosphates. Only 3'ATP underwent cation-catalyzed, nonenzymatic hydrolysis, with the primary product being 2':3'-cAMP. Because 3'-ADP and 3'-ATP are naturally occurring, this class of compounds may physiologically regulate adenylyl cyclases and possibly other enzymes, mediating responses that include a reduction in 3':5'-cAMP levels and consequent reductions in protein kinase A-activated pathways.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Inhibition of Adenylyl Cyclase by a Family of Newly Synthesized Adenine Nucleoside 3′-Polyphosphates

Désaubry

Shoshani

Johnson

1996

Journal of Biological Chemistry

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“…The identities of ATP, GTP and ppGpp were verified by comparison with commercial preparations of the nucleotides (Roche, TriLink) developed on the same TLC plate and visualized by UV shadowing. (p)ppGpp was prepared synthetically as described (Schattenkerk et al. , 1985), and its structure was verified by nuclear magnetic resonance and high‐resolution mass spectroscopy.…”

Section: Methodsmentioning

confidence: 99%

The identity of the transcription +1 position is crucial for changes in gene expression in response to amino acid starvation in Bacillus subtilis

Krásný

Tišerová

Jonák

et al. 2008

Molecular Microbiology

112

110

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SummaryWe identify here a pattern in the transcription start sites (+1A or +1G) of s A -dependent promoters of genes that are up-/downregulated in response to amino acid starvation (stringent response) in Bacillus subtilis. Upregulated promoters initiate mostly with ATP and downregulated promoters with GTP. These promoters appear to be sensitive to changes in initiating nucleoside triphosphate concentrations. During the stringent response in B. subtilis, when ATP and GTP levels change reciprocally, the identity of the +1 position (A or G) of these promoters is a factor important in their regulation. Mutations that change the identity of position +1 (A for G and vice versa) change the response of the promoter to amino acid starvation.

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“…S14 A and B). We chemically synthesized pure ppApp using a modification of the synthetic scheme that was originally developed for ppGpp (26) (SI Appendix, Fig. S15).…”

Section: Farel Toxicity Is Mediated By Accumulation Of Ppgpp and Ppappmentioning

confidence: 99%

A widespread toxin−antitoxin system exploiting growth control via alarmone signaling

Jimmy

Saha

Kurata

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

207

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Under stressful conditions, bacterial RelA-SpoT Homolog (RSH) enzymes synthesize the alarmone (p)ppGpp, a nucleotide second messenger. (p)ppGpp rewires bacterial transcription and metabolism to cope with stress, and, at high concentrations, inhibits the process of protein synthesis and bacterial growth to save and redirect resources until conditions improve. Single-domain small alarmone synthetases (SASs) are RSH family members that contain the (p)ppGpp synthesis (SYNTH) domain, but lack the hydrolysis (HD) domain and regulatory C-terminal domains of the long RSHs such as Rel, RelA, and SpoT. We asked whether analysis of the genomic context of SASs can indicate possible functional roles. Indeed, multiple SAS subfamilies are encoded in widespread conserved bicistronic operon architectures that are reminiscent of those typically seen in toxin−antitoxin (TA) operons. We have validated five of these SASs as being toxic (toxSASs), with neutralization by the protein products of six neighboring antitoxin genes. The toxicity of Cellulomonas marina toxSAS FaRel is mediated by the accumulation of alarmones ppGpp and ppApp, and an associated depletion of cellular guanosine triphosphate and adenosine triphosphate pools, and is counteracted by its HD domain-containing antitoxin. Thus, the ToxSAS–antiToxSAS system with its multiple different antitoxins exemplifies how ancient nucleotide-based signaling mechanisms can be repurposed as TA modules during evolution, potentially multiple times independently.

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Synthesis of riboguanosine pentaphosphate ppprGpp (Magic Spot II) via a phosphotriester approach

Cited by 17 publications

References 11 publications

Inhibition of Adenylyl Cyclase by a Family of Newly Synthesized Adenine Nucleoside 3′-Polyphosphates

Inhibition of Adenylyl Cyclase by a Family of Newly Synthesized Adenine Nucleoside 3′-Polyphosphates

The identity of the transcription +1 position is crucial for changes in gene expression in response to amino acid starvation in Bacillus subtilis

A widespread toxin−antitoxin system exploiting growth control via alarmone signaling

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