Author response: Flotillin-mediated membrane fluidity controls peptidoglycan synthesis and MreB movement

Zielińska, Aleksandra; Savietto, Abigail; Borges, Anabela; Martinez, Dênis; Berbon, Mélanie; Roelofsen, Joël R; Hartman, Alwin M.; Boer, Rinse de; Klei, Ida J. van der; Hirsch, A.; Habenstein, Birgit; Bramkamp, Marc; Scheffers, Dirk‐Jan

doi:10.7554/elife.57179.sa2

Cited by 3 publications

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In B. subtilis known flotillins are the integral membrane protein FloA and the peripheral FloT (Figure 4) (Dempwolff et al, 2012). However, a recent study has shown evidence that these proteins may be associated with fluid rather than rigid domains (Zielińska et al, 2020), challenging the established paradigm and questioning the use of flotillins as raft reporters in bacteria.…”

Section: Membrane Domainsmentioning

confidence: 99%

A How-To Guide for Mode of Action Analysis of Antimicrobial Peptides

Schäfer

Wenzel

2020

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Antimicrobial peptides (AMPs) are a promising alternative to classical antibiotics in the fight against multi-resistant bacteria. They are produced by organisms from all domains of life and constitute a nearly universal defense mechanism against infectious agents. No drug can be approved without information about its mechanism of action. In order to use them in a clinical setting, it is pivotal to understand how AMPs work. While many pore-forming AMPs are well-characterized in model membrane systems, non-pore-forming peptides are often poorly understood. Moreover, there is evidence that pore formation may not happen or not play a role in vivo. It is therefore imperative to study how AMPs interact with their targets in vivo and consequently kill microorganisms. This has been difficult in the past, since established methods did not provide much mechanistic detail. Especially, methods to study membrane-active compounds have been scarce. Recent advances, in particular in microscopy technology and cell biological labeling techniques, now allow studying mechanisms of AMPs in unprecedented detail. This review gives an overview of available in vivo methods to investigate the antibacterial mechanisms of AMPs. In addition to classical mode of action classification assays, we discuss global profiling techniques, such as genomic and proteomic approaches, as well as bacterial cytological profiling and other cell biological assays. We cover approaches to determine the effects of AMPs on cell morphology, outer membrane, cell wall, and inner membrane properties, cellular macromolecules, and protein targets. We particularly expand on methods to examine cytoplasmic membrane parameters, such as composition, thickness, organization, fluidity, potential, and the functionality of membrane-associated processes. This review aims to provide a guide for researchers, who seek a broad overview of the available methodology to study the mechanisms of AMPs in living bacteria.

show abstract

Section: Membrane Domainsmentioning

confidence: 99%

A How-To Guide for Mode of Action Analysis of Antimicrobial Peptides

Schäfer

Wenzel

2020

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

show abstract

The Bacterial Cytoskeleton Spatially Confines Functional Membrane Microdomains

Machta

et al. 2020

Preprint

View full text Add to dashboard Cite

22Cell membranes laterally segregate into microdomains enriched in certain lipids and scaffold 23 proteins. Membrane microdomains modulate protein-protein interactions and are essential for cell 24 polarity, signaling and membrane trafficking. How cells organize their membrane microdomains, and 25 the physiological importance of these microdomains, is unknown. In eukaryotes, the cortical actin 26 cytoskeleton is proposed to act like a fence, constraining the dynamics of membrane microdomains. 27Like their eukaryotic counterparts, bacterial cells have functional membrane microdomains (FMMs) 28 that act as platforms for the efficient oligomerization of protein complexes. In this work, we used the 29 model organism Bacillus subtilis to demonstrate that FMM organization and movement depend 30 primarily on the interaction of FMM scaffold proteins with the domains' protein cargo, rather than with 31 domain lipids. Additionally, the MreB actin-like cytoskeletal network that underlies the bacterial 32 membrane was found to frame areas of the membrane in which FMM mobility is concentrated. 33Variations in membrane fluidity did not affect FMM mobility whereas alterations in cell wall organization 34 affected FMM mobility substantially. Interference with MreB organization alleviates FMM spatial 35 confinement whereas, by contrast, inhibition of cell wall synthesis strengthens FMM confinement. The 36 restriction of FMM lateral mobility by the submembranous actin-like cytoskeleton or the extracellular 37 wall cytoskeleton appears to be a conserved mechanism in prokaryotic and eukaryotic cells for 38 localizing functional protein complexes in specific membrane regions, thus contributing to the 39 organization of cellular processes. 40 41 42 43 48 actually heterogeneous fluids, with different membrane lipid species laterally segregating into sub-49 micrometer domains, driven by their physico-chemical properties 2-4 . This heterogeneous organization 50 of membrane lipids leads to a heterogeneous distribution of the embedded membrane proteins, which 51 appears essential for their functionality 5 . Membrane microdomains are widely present, mobile 52 structures that vary in size from transient protein clusters measured in nm 6 to large micron-sized stable 53 domains such as desmosomes 7 . These domains harbor proteins specialized in membrane trafficking, 54 cell-cell adhesion, and signal transduction, and they provide targets for viral entry 8 . 56It is now appreciated that membrane proteins can significantly impact membrane organization, in 57 particular stabilizing membrane domains. For instance, the scaffold activity of the flotillin proteins FLO1 58 and FLO2 plays an important role in stabilizing some eukaryotic membrane rafts ("lipid rafts") 9-11 ; it 59 would seem that this raft organization occurs in part through the capture and stabilization of specific 60 lipids by flotillins 12,13 . In addition, the activity of the flotillin scaffold contributes to the recruitment of 61 other raft-associated proteins, thus facilitatin...

show abstract

Escherichia coli SPFH Membrane Microdomain Proteins HflKC Contribute to Aminoglycoside and Oxidative Stress Tolerance

et al. 2023

View full text Add to dashboard Cite

Eukaryotic cells often segregate physiological processes in cholesterol-rich functional membrane microdomains. These domains are also called lipid rafts and contain proteins of the s tomatin, p rohibitin, f lotillin, and H flK/C (SPFH) superfamily, which are also present in prokaryotes but have been mostly studied in Gram-positive bacteria.

show abstract

Author response: Flotillin-mediated membrane fluidity controls peptidoglycan synthesis and MreB movement

Cited by 3 publications

References 0 publications

A How-To Guide for Mode of Action Analysis of Antimicrobial Peptides

A How-To Guide for Mode of Action Analysis of Antimicrobial Peptides

The Bacterial Cytoskeleton Spatially Confines Functional Membrane Microdomains

Escherichia coli SPFH Membrane Microdomain Proteins HflKC Contribute to Aminoglycoside and Oxidative Stress Tolerance

Contact Info

Product

Resources

About