Dataset Information

Disassembly of Functional Membrane Microdomains Relapses Antibiotic Resistance in Staphylococcus aureus

ABSTRACT: Bacteria spatially confine cellular processes like protease secretion and signal transduction in membrane platforms termed functional membrane microdomains, which in certain organisational and functional features resemble the lipid rafts of eukaryotic cells. However, a rigorous understanding of their composition, assembly and biological significance is unknown. Here we use the human pathogen methicillin-resistant Staphylococcus aureus (MRSA) to show that the organization of these platforms requires a preferential interaction between unphosphorylated membrane saccharolipids and the scaffold protein flotillin. This interaction leads to their accumulation in specific membrane microdomains concomitantly to the attraction of membrane-associated multimeric complexes, for which flotillin promotes efficient oligomerization. One of these harbored proteins is the penicillin-binding protein PBP2a, responsible for penicillin resistance in MRSA. We took PBP2a as showcase to demonstrate that flotillin mutants are also defective in PBP2a oligomerization and activity. Thus, perturbation of microdomains assembly, using commercially available drugs, interferes with PBP2a oligomerization and causes a relapse of MRSA penicillin resistance in vitro and in vivo, resulting in MRSA infections susceptible to conventional penicillin treatments. Our study shows that bacterial cells organize sophisticated programs for cellular compartmentalization and unravels a novel strategy to develop antimicrobial therapies for multi-drug resistance pathogens.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Staphylococcus Aureus

SUBMITTER: Jens Vanselow

LAB HEAD: Andreas Schlosser

PROVIDER: PXD006546 | Pride | 2019-01-08

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	2016-03-08_Fusion_5502.raw	Raw
	2016-03-08_Fusion_5504.raw	Raw
	2016-03-08_Fusion_5506.raw	Raw
	2016-03-08_Fusion_5511.raw	Raw
	2016-03-08_Fusion_5513.raw	Raw

Items per page:

1 - 5 of 15

Publications

Membrane Microdomain Disassembly Inhibits MRSA Antibiotic Resistance.

García-Fernández Esther E Koch Gudrun G Wagner Rabea M RM Fekete Agnes A Stengel Stephanie T ST Schneider Johannes J Mielich-Süss Benjamin B Geibel Sebastian S Markert Sebastian M SM Stigloher Christian C Lopez Daniel D

Cell 20171102 6

A number of bacterial cell processes are confined functional membrane microdomains (FMMs), structurally and functionally similar to lipid rafts of eukaryotic cells. How bacteria organize these intricate platforms and what their biological significance is remain important questions. Using the pathogen methicillin-resistant Staphylococcus aureus (MRSA), we show here that membrane-carotenoid interaction with the scaffold protein flotillin leads to FMM formation, which can be visualized using super- ...[more]

PMID: 29103614

Similar Datasets

Project description:The simultaneous overexpression of flotillins 1 and 2 is observed in many cancers and considered as a marker of poor prognosis, notably in breast cancer. Flotillin-overexpression was shown to participate in the invasive behavior of cancer cell leading to metastasis. To better characterize the specific contribution of flotillin upregulation in the acquisition of cellular invasive properties, we stably expressed at high level exogenous flotillin 1 (HA-tagged) and flotillin 2 (mCherry tagged) in the non-tumoral mammary epithelial MCF10A human cells. The obtained cell line was named MCF10AF1F2. As a control, we generated in parallel the MCF10AmCherry cell line, stably expressing mCherry alone. We compared by RNA-sequencing (RNA-seq) the expression profiles of MCF10AmCh cells and MCF10AF1F2 cells. This allows to identify the gene expression changes and the downstream signaling pathways associated with flotillin upregulation. Using two differential gene expression analysis pipelines (Tuxedo and DSeq2), we identified 232 upregulated and 570 downregulated genes (802 genes in total, fold change 2.8, P<0.05) in MCF10AF1F2 cells compared with control cells. Gene ontology (GO) cluster analysis) revealed that flotillin upregulation was associated with deregulation of genes involved in extracellular matrix, cell adhesion, migration, differentiation, apoptosis and signaling pathways, particularly MAP kinases. Because these data suggested that MCF10AF1F2 cells underwent epithelio to mesenchymal transition (EMT), we compared the list of genes differentially in MCF10AF1F2 cells with the EMT transcriptional signature previously established by a meta-analysis (Gröger et al al. PloSOne 2012, https://doi.org/10.1371/journal.pone.0051136). One third (22/67) of the upregulated genes and more than half (39/62) of the downregulated genes in this EMT signature overlapped with the genes deregulated upon flotillin overexpression in MCF10A cells. Using complementary approaches, we confirmed that upregulation of flotillins un non-tumoral mammary epithelial cells is sufficient to promote EMT.