Synthesis and Uptake of the Compatible Solutes Ectoine and 5-Hydroxyectoine by Streptomyces coelicolor A3(2) in Response to Salt and Heat Stresses

ABSTRACT Streptomyces coelicolor A3(2) synthesizes ectoine and 5-hydroxyectoine upon the imposition of either salt (0.5 M NaCl) or heat stress (39°C). The cells produced the highest cellular levels of these compatible solutes when both stress conditions were simultaneously imposed. Protection against either severe salt (1.2 M NaCl) or heat stress (39°C) or a combination of both environmental cues could be accomplished by adding low concentrations (1 mM) of either ectoine or 5-hydroxyectoine to S. coelicolor A3(2) cultures. The best salt and heat stress protection was observed when a mixture of ectoine and 5-hydroxyectoine (0.5 mM each) was provided to the growth medium. Transport assays with radiolabeled ectoine demonstrated that uptake was triggered by either salt or heat stress. The most effective transport and accumulation of [14C]ectoine by S. coelicolor A3(2) were achieved when both environmental cues were simultaneously applied. Our results demonstrate that the accumulation of the compatible solutes ectoine and 5-hydroxyectoine allows S. coelicolor A3(2) to fend off the detrimental effects of both high salinity and high temperature on cell physiology. We also characterized the enzyme (EctD) required for the synthesis of 5-hydroxyectoine from ectoine, a hydroxylase of the superfamily of the non-heme-containing iron(II)- and 2-oxoglutarate-dependent dioxygenases (EC 1.14.11). The gene cluster (ectABCD) encoding the enzymes for ectoine and 5-hydroxyectoine biosynthesis can be found in the genome of S. coelicolor A3(2), Streptomyces avermitilis, Streptomyces griseus, Streptomyces scabiei, and Streptomyces chrysomallus, suggesting that these compatible solutes play an important role as stress protectants in the genus Streptomyces.

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Genome Sequence of the Streptomycin-Producing Microorganism Streptomyces griseus IFO 13350

Journal of Bacteriology ◽

10.1128/jb.00204-08 ◽

2008 ◽

Vol 190 (11) ◽

pp. 4050-4060 ◽

Cited By ~ 389

Author(s):

Yasuo Ohnishi ◽

Jun Ishikawa ◽

Hirofumi Hara ◽

Hirokazu Suzuki ◽

Miwa Ikenoya ◽

...

Keyword(s):

Genome Sequence ◽

Streptomyces Coelicolor ◽

Streptomyces Griseus ◽

Aminoglycoside Antibiotic ◽

Gene Clusters ◽

Open Reading Frames ◽

Streptomyces Avermitilis ◽

Trna Genes ◽

Biosynthesis Gene Cluster ◽

Regulatory Cascade

ABSTRACT We determined the complete genome sequence of Streptomyces griseus IFO 13350, a soil bacterium producing an antituberculosis agent, streptomycin, which is the first aminoglycoside antibiotic, discovered more than 60 years ago. The linear chromosome consists of 8,545,929 base pairs (bp), with an average G+C content of 72.2%, predicting 7,138 open reading frames, six rRNA operons (16S-23S-5S), and 66 tRNA genes. It contains extremely long terminal inverted repeats (TIRs) of 132,910 bp each. The telomere's nucleotide sequence and secondary structure, consisting of several palindromes with a loop sequence of 5′-GGA-3′, are different from those of typical telomeres conserved among other Streptomyces species. In accordance with the difference, the chromosome has pseudogenes for a conserved terminal protein (Tpg) and a telomere-associated protein (Tap), and a novel pair of Tpg and Tap proteins is instead encoded by the TIRs. Comparisons with the genomes of two related species, Streptomyces coelicolor A3(2) and Streptomyces avermitilis, clarified not only the characteristics of the S. griseus genome but also the existence of 24 Streptomyces-specific proteins. The S. griseus genome contains 34 gene clusters or genes for the biosynthesis of known or unknown secondary metabolites. Transcriptome analysis using a DNA microarray showed that at least four of these clusters, in addition to the streptomycin biosynthesis gene cluster, were activated directly or indirectly by AdpA, which is a central transcriptional activator for secondary metabolism and morphogenesis in the A-factor (a γ-butyrolactone signaling molecule) regulatory cascade in S. griseus.

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Oxidation and amidation of salicylate by Streptomyces species

Canadian Journal of Microbiology ◽

10.1139/m96-111 ◽

1996 ◽

Vol 42 (8) ◽

pp. 867-869 ◽

Cited By ~ 2

Author(s):

Stanislav Pospíšil ◽

Věra Přikrylová ◽

Jan Němeček ◽

Jaroslav Spížek

Keyword(s):

Time Course ◽

Streptomyces Coelicolor ◽

Streptomyces Griseus ◽

Culture Broth ◽

Streptomyces Avermitilis ◽

Resonance Spectroscopy ◽

Streptomyces Species ◽

Streptomyces Spp ◽

Streptomyces Cinnamonensis ◽

Layer Chromatography

Seven streptomycete strains were tested for biotransformation of salicylate. The products were identified by nuclear magnetic resonance spectroscopy and three types of conversion were found. Streptomyces cinnamonensis and Streptomyces spectabilis formed gentisate and salicylamide concurrently. Streptomyces rimosus transformed salicylate to salicylamide. Streptomyces lividans, Streptomyces coelicolor, Streptomyces griseus and Streptomyces avermitilis produced only gentisate. Time course studies of salicylate conversion by thin-layer chromatography and high pressure liquid chromatography showed that salicylamide was accumulated in the culture broth, whereas gentisate was further metabolized.Key words: salicylate, gentisate, salicylamide, biotransformation, Streptomyces spp.

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Oxidative/heat stress enhanced production of chitosanase from Streptomyces griseus cells through its interaction with liposome

Journal of Bioscience and Bioengineering ◽

10.1016/j.jbiosc.2009.06.010 ◽

2009 ◽

Vol 108 (6) ◽

pp. 471-476 ◽

Cited By ~ 3

Author(s):

Kien Xuan Ngo ◽

Hiroshi Umakoshi ◽

Haruyuki Ishii ◽

Huong Thi Bui ◽

Toshinori Shimanouchi ◽

...

Keyword(s):

Heat Stress ◽

Streptomyces Griseus ◽

Enhanced Production

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The Level of AdpA Directly Affects Expression of Developmental Genes in Streptomyces coelicolor

Journal of Bacteriology ◽

10.1128/jb.05734-11 ◽

2011 ◽

Vol 193 (22) ◽

pp. 6358-6365 ◽

Cited By ~ 29

Author(s):

Marcin Wolański ◽

Rafał Donczew ◽

Agnieszka Kois-Ostrowska ◽

Paweł Masiewicz ◽

Dagmara Jakimowicz ◽

...

Keyword(s):

Early Stage ◽

Response Regulator ◽

Streptomyces Coelicolor ◽

Streptomyces Griseus ◽

Morphological Differentiation ◽

Aerial Mycelium ◽

Content Type ◽

Protein Functions

AdpA is a key regulator of morphological differentiation inStreptomyces. In contrast toStreptomyces griseus, relatively little is known about AdpA protein functions inStreptomyces coelicolor. Here, we report for the first time the translation accumulation profile of theS. coelicoloradpA(adpASc) gene; the level ofS. coelicolorAdpA (AdpASc) increased, reaching a maximum in the early stage of aerial mycelium formation (after 36 h), and remained relatively stable for the next several hours (48 to 60 h), and then the signal intensity decreased considerably. AdpAScspecifically binds theadpAScpromoter regionin vitroandin vivo, suggesting that its expression is autoregulated; surprisingly, in contrast toS. griseus, the protein presumably acts as a transcriptional activator. We also demonstrate a direct influence of AdpAScon the expression of several genes whose products play key roles in the differentiation ofS. coelicolor: STI, a protease inhibitor; RamR, an atypical response regulator that itself activates expression of the genes for a small modified peptide that is required for aerial growth; and ClpP1, an ATP-dependent protease. The diverse influence of AdpAScprotein on the expression of the analyzed genes presumably results mainly from different affinities of AdpAScprotein to individual promoters.

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Identification and Gene Disruption of Small Noncoding RNAs in Streptomyces griseus

Journal of Bacteriology ◽

10.1128/jb.00087-09 ◽

2009 ◽

Vol 191 (15) ◽

pp. 4896-4904 ◽

Cited By ~ 18

Author(s):

Takeaki Tezuka ◽

Hirofumi Hara ◽

Yasuo Ohnishi ◽

Sueharu Horinouchi

Keyword(s):

Type Strain ◽

Wild Type Strain ◽

Noncoding Rnas ◽

Streptomyces Griseus ◽

Streptomyces Avermitilis ◽

Dependent Manner ◽

Wild Type ◽

Srna Candidate ◽

Small Noncoding Rnas ◽

Nuclease Mapping

ABSTRACT Small noncoding RNAs (sRNAs) have been shown to control diverse cellular processes in prokaryotes. To identify and characterize novel bacterial sRNAs, a gram-positive, soil-inhabiting, filamentous bacterium, Streptomyces griseus, was examined, on the assumption that Streptomyces should express sRNAs as important regulators of morphological and physiological differentiation. By bioinformatics investigation, 54 sRNA candidates, which were encoded on intergenic regions of the S. griseus chromosome and were highly conserved in those of both Streptomyces coelicolor A3(2) and Streptomyces avermitilis, were selected. Of these 54 sRNA candidates, 17 transcripts were detected by Northern blot analysis of the total RNAs isolated from cells grown on solid medium. Then, the direction of transcription of each sRNA candidate gene was determined by S1 nuclease mapping, followed by exclusion of four sRNA candidates that were considered riboswitches of their downstream open reading frames (ORFs). Finally, a further sRNA candidate was excluded because it was cotranscribed with the upstream ORF determined by reverse transcription-PCR. Thus, 12 sRNAs ranging in size from 40 to 300 nucleotides were identified in S. griseus. Seven of them were apparently transcribed in a growth phase-dependent manner. Furthermore, of the 12 sRNAs, the expression profiles of 7 were significantly influenced by a mutation of adpA, which encodes the central transcriptional regulator of the A-factor regulatory cascade involved in both morphological differentiation and secondary metabolism in S. griseus. However, disruption of all 12 sRNA genes showed no detectable phenotypic changes; all the disruptants grew and formed aerial mycelium and spores with the same time course as the wild-type strain on various media and produced streptomycin similarly to the wild-type strain.

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Mannosylglycerate and Di-myo-Inositol Phosphate Have Interchangeable Roles during Adaptation of Pyrococcus furiosus to Heat Stress

Applied and Environmental Microbiology ◽

10.1128/aem.00559-14 ◽

2014 ◽

Vol 80 (14) ◽

pp. 4226-4233 ◽

Cited By ~ 16

Author(s):

Ana M. Esteves ◽

Sanjeev K. Chandrayan ◽

Patrick M. McTernan ◽

Nuno Borges ◽

Michael W. W. Adams ◽

...

Keyword(s):

Heat Stress ◽

Pyrococcus Furiosus ◽

Inositol Phosphate ◽

Compatible Solutes ◽

Optimal Growth ◽

Growth Conditions ◽

Cell Protection ◽

Content Type ◽

Molecular Events ◽

Supraoptimal Temperatures

ABSTRACTMarine hyperthermophiles accumulate small organic compounds, known as compatible solutes, in response to supraoptimal temperatures or salinities.Pyrococcus furiosusis a hyperthermophilic archaeon that grows optimally at temperatures near 100°C. This organism accumulates mannosylglycerate (MG) and di-myo-inositol phosphate (DIP) in response to osmotic and heat stress, respectively. It has been assumed that MG and DIP are involved in cell protection; however, firm evidence for the roles of these solutes in stress adaptation is still missing, largely due to the lack of genetic tools to produce suitable mutants of hyperthermophiles. Recently, such tools were developed forP. furiosus, making this organism an ideal target for that purpose. In this work, genes coding for the synthases in the biosynthetic pathways of MG and DIP were deleted by double-crossover homologous recombination. The growth profiles and solute patterns of the two mutants and the parent strain were investigated under optimal growth conditions and also at supraoptimal temperatures and NaCl concentrations. DIP was a suitable replacement for MG during heat stress, but substitution of MG for DIP and aspartate led to less efficient growth under conditions of osmotic stress. The results suggest that the cascade of molecular events leading to MG synthesis is tuned for osmotic adjustment, while the machinery for induction of DIP synthesis responds to either stress agent. MG protects cells against heat as effectively as DIP, despite the finding that the amount of DIP consistently increases in response to heat stress in the nine (hyper)thermophiles examined thus far.

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The Product of a Developmental Gene, crgA, That Coordinates Reproductive Growth in Streptomyces Belongs to a Novel Family of Small Actinomycete-Specific Proteins

Journal of Bacteriology ◽

10.1128/jb.185.22.6678-6685.2003 ◽

2003 ◽

Vol 185 (22) ◽

pp. 6678-6685 ◽

Cited By ~ 13

Author(s):

Ricardo Del Sol ◽

Andrew Pitman ◽

Paul Herron ◽

Paul Dyson

Keyword(s):

Antibiotic Production ◽

Streptomyces Coelicolor ◽

Orthologous Gene ◽

Streptomyces Avermitilis ◽

Antibiotic Biosynthesis ◽

Reproductive Growth ◽

Solid Media ◽

Aerial Hyphae ◽

A Chain ◽

Membrane Spanning

ABSTRACT On solid media, the reproductive growth of Streptomyces involves antibiotic biosynthesis coincident with the erection of filamentous aerial hyphae. Following cessation of growth of an aerial hypha, multiple septation occurs at the tip to form a chain of unigenomic spores. A gene, crgA, that coordinates several aspects of this reproductive growth is described. The gene product is representative of a well-conserved family of small actinomycete proteins with two C-terminal hydrophobic-potential membrane-spanning segments. In Streptomyces avermitilis, crgA is required for sporulation, and inactivation of the gene abolished most sporulation septation in aerial hyphae. Disruption of the orthologous gene in Streptomyces coelicolor indicates that whereas CrgA is not essential for sporulation in this species, during growth on glucose-containing media, it influences the timing of the onset of reproductive growth, with precocious erection of aerial hyphae and antibiotic production by the mutant. Moreover, CrgA subsequently acts to inhibit sporulation septation prior to growth arrest of aerial hyphae. Overexpression of CrgA in S. coelicolor, uncoupling any nutritional and growth phase-dependent regulation, results in growth of nonseptated aerial hyphae on all media tested, consistent with a role for the protein in inhibiting sporulation septation.

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