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Effect of myricetin treatment on Roseovarius tolerans EL-164 during exponential and early stationary phase growth.

ABSTRACT: Discovery of cryptic natural products can be expedited by the use of high throughput elicitor screening (HiTES). In this study, we discover that catechol-containing flavonoids elicit the production of a new group of ceramide lipids in the marine bacterium Roseovarius tolerans EL-164. To investigate the mechanism of elicitation of myricetin, a model catechol flavonoid, we used RNA-seq to determine the molecular response. The transcriptomic dataset illustrated that iron-starvation genes, as well as the import and catabolism of carbon sources, specifically branched chain amino acids, were upregulated upon myricetin treatment. Simultaneously, complex IV of the electron transport chain was universally downregulated as were oxidative stress responses. Together, along with further follow up studies, we were able to support the model that myricetin reduces the bioavailability of iron and reduced oxidative phosphorylation and the proton motive force. Our data suggests that the hampering of the proton motive force is what initiates the upregulation of the ceramide lipids.

ORGANISM(S): Roseovarius tolerans

PROVIDER: GSE279554 | GEO | 2024/10/21

REPOSITORIES: GEO

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Project description:Abstract: The crenarchaeal order Sulfolobales collectively contains at least five major terminal oxidase complexes. Based on genome sequence information, all five complexes are found only in Metallosphaera sedula and Sulfolobus tokodaii, the two sequenced Sulfolobales capable of iron oxidization. While specific respiratory complexes in certain Sulfolobales have been characterized previously as proton pumps for maintaining intracellular pH and generating proton motive force (pmf), their contribution to sulfur and iron biooxidation has not been considered. For M. sedula growing in the presence of ferrous iron and reduced inorganic sulfur compounds (RISCs), global transcriptional analysis was used to track the response of specific genes associated with these complexes, as well as other known and putative respiratory electron transport chain elements. ORFs from all five terminal oxidase or bc1-like complexes were stimulated on one or more conditions tested. Components of the fox (Msed0467-0489) and soxNL-cbsABA (Msed0500-0505) terminal/quinol oxidase clusters were triggered by ferrous iron, while the soxABCDD' terminal oxidase cluster (Msed0285-0291) were induced by tetrathionate and S°. Chemolithotrophic electron transport elements, including a putative tetrathionate hydrolase (Msed0804), a novel polysulfide/sulfur/DMSO reductase-like complex (Msed0812-0818), and a novel heterodisulfide reductase-like complex (Msed1542-1550), were also stimulated by RISCs. Furthermore, several hypothetical proteins were found to have strong responses to ferrous iron or RISCs, suggesting additional candidates in iron or sulfur oxidation-related pathways. From this analysis, a comprehensive model for electron transport in M. sedula could be proposed as the basis for examining specific details of iron and sulfur oxidation in this bioleaching archaeon. 5-slide loop of Mse cells includes 5 conditions tested: yeast exact (Y), yeast extract + ferrous sulfate (YFS), yeast extract + potassium sulfate (YKS), yeast extract + potassium tetrathionate (YKT), and yeast extract + elemental sulfur (YS). Half of an RNA sample for one condition was labeled with Cy3 while the other half was labeled with Cy5. The two differently labeled samples were run on different slides. Each probe is spotted on each slide 5 times (5 replicates; spot intensities for all replicates on slide provided in associated raw data file).

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Effect of myricetin treatment on Roseovarius tolerans EL-164 during exponential and early stationary phase growth.

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