Radiolabeling of Treponema pallidum (Nichols virulent strain) in vitro with precursors for protein and RNA biosynthesis

Sandok, P L; Jenkin, Howard M.

doi:10.1128/iai.22.1.22-28.1978

Cited by 6 publications

(4 citation statements)

References 23 publications

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“…Analysis of acid-hydrolyzed nucleic acid showed that glucose was utilized for the synthesis of both ribose and deoxyribose, which implicates the action of several enzymes for the utilization of pentoses in the biosynthesis of nucleotides (28). These findings were consistent with the ability of T. pallidum to incorporate nitrogenous bases into nucleic acid (32,33,36). Also noteworthy were the investigations by Baseman and coworkers (8,32) which showed that nucleosides were also utilized for nucleic acid biosynthesis.…”

Section: Discussionsupporting

confidence: 74%

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Distribution of glucose incorporated into macromolecular material by treponema pallidum

Barbieri¹,

Austin²,

Cox³

1981

Infect Immun

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Treponema pallidum was observed to incorporate glucose carbons into lipids, ribonucleic acid, deoxyribonucleic acid, and protein. Only the glycerol portions of phosphatidylcholine and phosphatidylglycerol contained glucose-derived carbons. Incorporation of exogenous choline into phosphatidylcholine was detected. Glucose was incorporated into only the pentoses of nucleic acids. About 50% of the glucose incorporated into protein was present in only one amino acid, aspartate. Evidence suggests that aspartate synthesis could follow the conversion of phosphoenolpyruvate to oxalacetic acid by a guanosine 5'-diphosphate-dependent phosphoenolpyruvate carboxykinase. Treponemapallidum degrades glucose via the Embden-Meyerhof and hexose monophosphate pathways (37), producing C02, acetate, pyruvate, and lactate (7). Nicotinamide adenine dinucleotide (NAD)-dependent lactate dehydrogenase catalyzes the reduction of pyruvate to lactate (37), whereas an oxidase, similar to the pyruvate oxidase of Lactobacillus delbruckii, catalyzes the oxidation of pyruvate to C02 and acetylphosphate (4). Glucose catabolism appears to be coupled to energy generation through both substrate-level (4, 37) and oxidative phosphorylation (26). Recently, T. pallidum has been shown to incorporate glucose into trichloroacetic acid-precipitable material (3). The purpose of this study was to determine the fate of glucose carbons incorporated into macromolecular components. MATERIALS AND METHODS Preparation of treponeme suspensions. Propagation, extraction, purification, and counting of the Nichols strain of virulent T. pallidum have been described (4, 26, 37). Extraction of treponemes from infected testicles was performed for 1 h at room temperature in a maintenance medium (BME) containing 34.0 mM NaCl, 5.4 mM KCI, 0.4 mM MgSO4, 0.34 mM CaCl12, 10.0 mM MOPS (morpholinopropanesulfonic acid), 10.0 mM TES [N-tris(hydroxymethyl)methyl-2aminoethanesulfonic acid], 15 mM HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), 3.0 ,M thiamine hydrochloride, 0.015 ,uM vitamin B12, 0.4 ,M biotin, 0.4 ,uM p-amino-benzoic acid, 0.2 AM nicotinamide, 0.1 MM pyridoxine hydrochloride, 0.2 ,M folic acid, 4.5 AM calcium pantothenate, 10.0 yM Na2HPO4, 0.08 mM ornithine, 0.02 mM NAD, 16.0 mM NaHCO3, 0.2% bovine serum albumin, and 10.0 mM reduced glutathione, at a final pH of 7.4. Testicular suspensions were shaken under atmospheric 02 tensions for 1 h at room temperature and 1 h at 4°C.

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Section: Discussionsupporting

confidence: 74%

“…Studies on the anabolic capacities of T. pallidum have described the incorporation of amino acids into proteins (5, 6) and nucleosides and nitrogenous bases into nucleic acids (8,32,36). The treponeme has also been observed to sequester free fatty acids from the environment (37).…”

Section: Discussionmentioning

confidence: 99%

Distribution of glucose incorporated into macromolecular material by treponema pallidum

Barbieri¹,

Austin²,

Cox³

1981

Infect Immun

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“…pallidum was shown by Sandok and Jenkin in 1978 [46]. This study confirmed the absorption of serine, amino acid hydrolysates and uracil by T. pallidum in vitro by radiolabeling [46].…”

Section: Media Tested In the Attempts To Culture T Pallidumsupporting

confidence: 81%

“…Conversely, Noguchi strains T. refringens [11] Nichols avirulent T. refringens [11] in co-culture assays with mammalian cells, the positive impact of amino acids on the survival of T. pallidum was shown by Sandok and Jenkin in 1978 [46]. This study confirmed the absorption of serine, amino acid hydrolysates and uracil by T. pallidum in vitro by radiolabeling [46].…”

Section: Use Of Amino Acids As Nutrientssupporting

confidence: 67%

A review of in vitro attempts to develop the axenic culture of Treponema pallidum and genomics-based suggestions to achieve this elusive goal

Belkacemi

Alou

Khelaifia

et al. 2021

Journal of Medical Microbiology

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To date, the axenic culture of Treponema pallidum remains a challenge in the field of microbiology despite countless attempts. Here, we conducted a comprehensive bibliographic analysis using several databases and search engines, namely Pubmed, Google scholar, Google, Web of Science and Scopus. Numerous unsuccessful empiric studies have been conducted and evaluated using as criteria dark-field microscopic observation of motile spiral shaped cells in the culture and virulence of the culture through rabbit infectivity. All of these studies failed to induce rabbit infectivity, even when deemed positive after microscopic observation leading to the misnomer of avirulent T. pallidum . In fact, this criterion was improperly chosen because not all spiral shaped cells are T. pallidum . However, these studies led to the formulation of culture media particularly favourable to the growth of several species of Treponema, including Oral Microbiology and Immunology, Zürich medium (OMIZ), Oral Treponeme Enrichment Broth (OTEB) and T-Raoult, thus allowing the increase in the number of cultivable strains of Treponema . The predicted metabolic capacities of T. pallidum show limited metabolism, also exhibited by other non-cultured and pathogenic Treponema species, in contrast to cultured Treponema species. The advent of next generation sequencing represents a turning point in this field, as the knowledge inferred from the genome can finally lead to the axenic culture of T. pallidum .

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Restriction analysis of DNA from Treponema pallidum, the causative agent of syphilis

Nath

1983

Molec Gen Genet

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When purified DNA from pathogenic Treponema pallidum is digested with restriction endonucleases it results in the formation of discrete DNA fragments which range between 2.5 to 10 Kilobase pairs. No such precise fragmentation occurs with DNA isolated from nonpathogenic T. pallidum. The appearance of the discrete restriction fragments from the pathogenic T. pallidum DNA does not represent a contamination of satellite DNA from rabbit, the host in which the organism was propagated, but rather represents the presence of redundant DNA or DNA of less complexity in the pathogenic T. pallidum DNA preparation.

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Radiolabeling of Treponema pallidum (Nichols virulent strain) in vitro with precursors for protein and RNA biosynthesis

Cited by 6 publications

References 23 publications

Distribution of glucose incorporated into macromolecular material by treponema pallidum

Distribution of glucose incorporated into macromolecular material by treponema pallidum

A review of in vitro attempts to develop the axenic culture of Treponema pallidum and genomics-based suggestions to achieve this elusive goal

Restriction analysis of DNA from Treponema pallidum, the causative agent of syphilis

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