Characterization and Persistence of Actinophage RP2 Isolated from Streptomyces rimosus ATCC 10970

Hranueli, Daslav; Pigac, Jasenka; Vešligaj, Margareta

doi:10.1099/00221287-114-2-295

Cited by 21 publications

(31 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The strain was maintained and grown in fermentation medium as described previously (35,36). Streptomyces plasmid pPFZ46 (26) was used as a source of the otcC gene and flanking sequences.…”

Section: Methodsmentioning

confidence: 99%

Ablation of the otcC Gene Encoding a Post-polyketide Hydroxylase from the Oxytetracyline Biosynthetic Pathway in Streptomyces rimosus Results in Novel Polyketides with Altered Chain Length

Perić-Concha

Borovička

Long

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC "minimal" polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex. Oxytetracycline (OTC)4 and related tetracycline polyketide compounds are potent inhibitors of bacterial protein synthesis displaying broad-spectrum activity against both Gram-positive and Gram-negative pathogens. Although the clinical use of the tetracyclines has declined in recent years due to the emergence of resistance in these bacteria, OTC is still the first drug of choice for the treatment of intracellular infections caused by Rickettsia, Chlamydia, and mycoplasma in penicillin-sensitive patients incapable of tolerating macrolides (1). Glycylcyclines are a new class of semi-synthetic tetracycline derivatives that exemplify how modification of a known antibiotic can be a successful strategy to obtain new compounds effective against pan-drug-resistant strains (2). To this end we have been elucidating the biosynthetic pathway leading to OTC in the producing actinomycete Streptomyces rimosus, as a potential route to clinically useful OTC products by combinatorial biosynthesis (3).Polyketide synthases (PKSs) catalyzing the formation of aromatic products such as OTC are dissociable enzyme complexes of largely monofunctional proteins that follow a mechanistic pathway similar to fatty acid biosynthesis (4). The synthesis of aromatic Type II polyketides usually begins with the condensation of an acetate unit derived from the decarboxylation of a malonyl-S-acyl carrier protein (ACP) to a malonyl-S-ACP extender unit. This reaction is catalyzed by a heterodimeric ketosynthase chain length factor (KS␣-KS␤) (5, 6). Preference for nonacetate primers has also been recognized in a number of aromatic PKSs including OTC, although presumably decarboxylative chain initiation catalyzed by the heterodimer can still occur in the absence of these non-acetate starter units (7). Manipulating the mechanism of non-acetate priming has lead to biosynthesis of novel polyketides (8, 9). The active site of the KS␣ also catalyzes iterative elongati...

show abstract

“…The strain was maintained and grown in fermentation medium as described previously (35,36). Streptomyces plasmid pPFZ46 (26) was used as a source of the otcC gene and flanking sequences.…”

Section: Methodsmentioning

confidence: 99%

Ablation of the otcC Gene Encoding a Post-polyketide Hydroxylase from the Oxytetracyline Biosynthetic Pathway in Streptomyces rimosus Results in Novel Polyketides with Altered Chain Length

Perić-Concha

Borovička

Long

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Although S. rimosus R6 is lysogenic for two prophages, RP2 and RP3 (104,193), this does not seem to cause any problems in antibiotic production.…”

Section: Localized Mutagenesismentioning

confidence: 99%

“…Hranueli and collaborators (104,106,107) detected and isolated free phage particles from a liquid culture of S. rimosus R7. The phage, designated RP2, appeared to be a typical temperate DNA phage producing turbid plaques in lawns of sensitive S. rimosus R6 cells.…”

Section: Prophagesmentioning

confidence: 99%

“…Treatment with UV light and other agents did not lead to induction of the prophage. Clear plaque mutants of RP2, which failed to lysogenize sensitive cultures, arose at a frequency of about 2 ϫ 10 Ϫ5 per phage particle (104,106,107). However, no virulent mutants that could infect lysogenic strains were observed, probably because multiple mutations would be necessary.…”

Section: Prophagesmentioning

confidence: 99%

“…In the early 1970s, studies following three avenues of research were initiated. These were the biochemical characterization of non-OTC-producing mutants and mapping of the OTC genes (180,200), characterization and persistence of S. rimosus phages (104,107,193,225), and genetic and physical characterization of circular and linear S. rimosus plasmids (31,41,72,80,201,212).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Genetics ofStreptomyces rimosus, the Oxytetracycline Producer

Petković

Cullum

Hranueli

et al. 2006

Microbiol Mol Biol Rev

101

View full text Add to dashboard Cite

SUMMARY From a genetic standpoint, Streptomyces rimosus is arguably the best-characterized industrial streptomycete as the producer of oxytetracycline and other tetracycline antibiotics. Although resistance to these antibiotics has reduced their clinical use in recent years, tetracyclines have an increasing role in the treatment of emerging infections and noninfective diseases. Procedures for in vivo and in vitro genetic manipulations in S. rimosus have been developed since the 1950s and applied to study the genetic instability of S. rimosus strains and for the molecular cloning and characterization of genes involved in oxytetracycline biosynthesis. Recent advances in the methodology of genome sequencing bring the realistic prospect of obtaining the genome sequence of S. rimosus in the near term.

show abstract

Molecular Biology Methods in Streptomyces rimosus, a Producer of Oxytetracycline

Slemc¹,

Pikl²,

Petković³

et al. 2021

Methods in Molecular Biology

View full text Add to dashboard Cite

Characterization and Persistence of Actinophage RP2 Isolated from Streptomyces rimosus ATCC 10970

Cited by 21 publications

References 18 publications

Ablation of the otcC Gene Encoding a Post-polyketide Hydroxylase from the Oxytetracyline Biosynthetic Pathway in Streptomyces rimosus Results in Novel Polyketides with Altered Chain Length

Ablation of the otcC Gene Encoding a Post-polyketide Hydroxylase from the Oxytetracyline Biosynthetic Pathway in Streptomyces rimosus Results in Novel Polyketides with Altered Chain Length

Genetics ofStreptomyces rimosus, the Oxytetracycline Producer

Molecular Biology Methods in Streptomyces rimosus, a Producer of Oxytetracycline

Contact Info

Product

Resources

About