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Proteome of the cellulose degradation system of Cytophaga hutchinsonii and Sporocytophaga myxococcoides

ABSTRACT: Both C. hutchinsonii and S. myxococcoides were grown on filter paper. Time points were taken in both early and late stages of growth to assess the cellulolytic proteins utilized during growth. As a non-cellulose control, the organisms were grown on pectin as sole carbon source. Further, the cells were also fractionated to seperate the cellular fractions in order to assess the localization of the proteins within the growing cells.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Cytophaga Hutchinsonii Sporocytophaga Myxococcoides

SUBMITTER: Johan Larsbrink

LAB HEAD: Johan Larsbrink

PROVIDER: PXD010836 | Pride | 2018-11-06

REPOSITORIES: pride

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Project description:Objectives: Carbapenem-resistant Acinetobacter baumannii (CRAB) are one of the most difficult pathogen to treat. The only drug recently approved by the FDA that is active against CRAB is cefiderocol. However, recent studies have shown higher all-cause mortality rate in the group of patients treated with cefiderocol, that may be due to heteroresistance, a phenotype characterized by the survival of a small proportion of cells in a population seemingly isogenic. Previous studies showed that adding human fluids to CRAB cultures can lead to CFDC heteroresistance. To better understand the nature of this phenomenon, we carried out molecular and phenotypic analyses of CRAB heteroresistant bacterial subpopulations. Methods: The CRAB strain AMA40 was cultured in the presence of cefiderocol and human pleural fluid (HPF) to isolate heteroresistant variants. Two of them, AMA40 IHC_1 and IHC_2, were subjected to whole genome sequencing and transcriptomic analysis to identify the mutations and transcriptomic changes responsible for the development of cefiderocol resistance. The impact of mutations on the pharmacodynamic activity of cefiderocol was assessed by susceptibility testing, EDTA and Boronic acid inhibition analysis, biofilm formation, and static time-kill assays. Results: Variants AMA40 IHC_1 and IHC_2 had 53 mutations, forty of which were common to both heteroresistant strains. None of the mutations are located inside genes associated with iron-uptake systems or β-lactam resistance. However, pipA, a gene associated with iron homeostasis in other species, was mutated in heteroresistant strains. Transcriptomic analyses revealed modifications in levels of expression of genes associated with antibiotic resistance. The blaNDM-1, blaADC-2, pbp3, and pbp1 were expressed at higher levels. At the same time, the carO and ompA genes’ expression was reduced. Collateral resistance to amikacin was observed in the heteroresistant variants. Static time-kill assays showed that when CA-MHB was supplemented with human serum albumin, the main protein component of HPF, cefiderocol killing activity was considerably reduced in all three strains. Conclusions: We conclude that heteroresistance to cefiderocol in CRAB, when exposed to fluids containing high HSA, is caused by mutations and modifications in the expression of genes associated with resistance to β-lactams.

			Action	DRS
	170530_ChEPC_A1.raw	Raw
	170530_ChEPC_A2.raw	Raw
	170530_ChEPC_B1.raw	Raw
	170530_ChEPC_B2.raw	Raw
	170530_ChEPC_C1.raw	Raw

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Proteome of the cellulose degradation system of Cytophaga hutchinsonii and Sporocytophaga myxococcoides

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