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ABSTRACT: Devosia lucknowensis L15

PROVIDER: PRJEB20274 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	FXWK01.dat.gz	Other
	FXWK01.fasta.gz	Fasta.gz
	FXWK01.master.dat	Other
	FXWK01.dat.gz	Other

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Devosia lucknowensis strain:L15 | isolate:DSM 25398

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Screening privileged alkylguanidinium motifs under host-mimicking conditions reveals a novel antibiotic with an unconventional mode of action

Project description:Screening large molecule libraries against pathogenic bacteria is often challenged by a low hit rate due to limited uptake, underrepresentation of antibiotic structural motifs and assays which do not resemble the infection conditions. To address these limitations, we here screen a library focused on alkylguanidinium compounds, a structural motif associated with antibiotic activity and enhanced uptake, under host-mimicking infection conditions against a panel of disease-associated bacteria. Several hit molecules were obtained with activities against Gram-positive as well as Gram-negative bacteria highlighting the fidelity of the general concept. Based on the best antibiotic activity, we selected one compound (L15) for in-depth mode of action studies. L15 exhibited bactericidal activity against Staphylococcus aureus USA300 Lac (JE2) with a minimum inhibitory concentration of 1.5 µM whereby a structure-activity relationship study with 18 derivatives proved the necessity of the guanidinium motif for antibiotic activity. Electron microscopy studies and depolarization assays at high L15 concentrations showed an impact on the cell membrane and integrity which was absent at the MIC concentration. Sequencing of L15-resistant strains revealed a mutation in an efflux pump but did not provide hints for direct targets. We thus performed affinity-based protein profiling with a L15 probe and identified signal peptidase IB (SpsB) as the most promising hit. Validation by activity assays, binding site identification and molecular docking demonstrated SpsB overactivation by L15, similar to a previously reported activator molecule that dysregulates protein secretion of this essential enzyme. Overall, this study highlights the need for unconventional screening strategies to identify novel antibiotics.

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Metabolite and transcript profiling of VvERF045 OE transgenic lines elucidate the role of this transcription factor in the grapevine ripening physiology

Project description:We report the transcriptomic profile of L15, a transgenic line overexpressing VvERF045, and the WT as a reference. The main aim is to understand the function of this transcription factor in the overall plant though the differences in the transcript abundance (RNA-seq) and metabolite production. Total RNA from L15 and WT was sequenced with an average genome coverage of more than 100X. Transcripts representing very different biological processes were grouped into ten major categories: protein phosphorilation, wax biosynthesis process, response to andogenous stimulus, cotyledonal vascular tissue/pattern formation, drug transmembrane transport stress, jasmonic acid biosynthesis, response to salt stress, defense response to bacterium, negative regulation. This transcription factor regulates genes involved in many different responses such as drought, sanility and defense suggesting it covers a key role into the plant, moreover into those processes also typical in ripening.

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