A Ubiquitous Platform for Bacterial Nanotube Biogenesis

Bhattacharya, Sumitra; Baidya, Amit K.; Pal, Ritesh Ranjan; Mamou, Gideon; Gatt, Yair E.; Margalit, Hanah; Rosenshine, Ilan; Ben-Yehuda, Sigal

doi:10.1016/j.celrep.2019.02.055

Cited by 41 publications

(66 citation statements)

References 47 publications

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“…6B, C). Furthermore, cells harboring a YFP fusion to an unrelated surface exposed membrane protein yielded a scattered pattern, which did not coincide with nanotubes 12,29 ( Supplementary Fig. 6D).…”

Section: Resultsmentioning

confidence: 99%

“…Molecular exchange assay. For detecting molecular exchange, antibiotic transfer assay was carried out as described previously 3,12 and in the legend to Supplementary Fig. 2.…”

Section: Methodsmentioning

confidence: 99%

“…CORE-dependent nanotube production was shown to be conserved among distinct species, signifying nanotube as a ubiquitous bacterial organelle. In accord, COREs from diverse species could restore nanotube generation and functionality in bacteria lacking endogenous CORE 9,12 .…”

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confidence: 93%

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Donor-delivered cell wall hydrolases facilitate nanotube penetration into recipient bacteria

2020

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Bacteria can produce membranous nanotubes that mediate contact-dependent exchange of molecules among bacterial cells. However, it is unclear how nanotubes cross the cell wall to emerge from the donor or to penetrate into the recipient cell. Here, we report that Bacillus subtilis utilizes cell wall remodeling enzymes, the LytC amidase and its enhancer LytB, for efficient nanotube extrusion and penetration. Nanotube production is reduced in a lytBC mutant, and the few nanotubes formed appear deficient in penetrating into target cells. Donor-derived LytB molecules localize along nanotubes and on the surface of nanotubeconnected neighbouring cells, primarily at sites of nanotube penetration. Furthermore, LytB from donor B. subtilis can activate LytC of recipient bacteria from diverse species, facilitating cell wall hydrolysis to establish nanotube connection. Our data provide a mechanistic view of how intercellular connecting devices can be formed among neighbouring bacteria.

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Section: Resultsmentioning

confidence: 99%

“…Molecular exchange assay. For detecting molecular exchange, antibiotic transfer assay was carried out as described previously 3,12 and in the legend to Supplementary Fig. 2.…”

Section: Methodsmentioning

confidence: 99%

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Donor-delivered cell wall hydrolases facilitate nanotube penetration into recipient bacteria

2020

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“…Furthermore, the helical structure of the export gate makes it likely that it is responsible for nucleating the helical filaments that assemble above it 11 . Interestingly, the EA has also recently been proposed to facilitate inward transport across the inner membrane associated with nanotubes 12,13 . The final component of the EA, FlhB/ SctU, has long been known to be essential for all T3SS-mediated protein secretion.…”

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confidence: 99%

The substrate specificity switch FlhB assembles onto the export gate to regulate type three secretion

et al. 2020

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Protein secretion through type-three secretion systems (T3SS) is critical for motility and virulence of many bacteria. Proteins are transported through an export gate containing three proteins (FliPQR in flagella, SctRST in virulence systems). A fourth essential T3SS protein (FlhB/SctU) functions to "switch" secretion substrate specificity once the growing hook/ needle reach their determined length. Here, we present the cryo-electron microscopy structure of an export gate containing the switch protein from a Vibrio flagellar system at 3.2 Å resolution. The structure reveals that FlhB/SctU extends the helical export gate with its four predicted transmembrane helices wrapped around FliPQR/SctRST. The unusual topology of the FlhB/SctU helices creates a loop wrapped around the bottom of the closed export gate. Structure-informed mutagenesis suggests that this loop is critical in gating secretion and we propose that a series of conformational changes in the T3SS trigger opening of the gate through interactions between FlhB/SctU and FliPQR/SctRST.

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“…OMV formation is ubiquitous and has many documented functions 9 . OMEs are less commonly observed, remain attached to the cell, and various morphologies can be seen extending from single cells including Myxococcus xanthus 14,15 , flavobacterium strain Hel3_A1_48 8 , Vibrio vulnificus 10 , Francisella novicida 28 , Shewanella oneidensis 7,29–31 , and as cell-cell connections in Bacillus subtilis 32–34 and Eschericia coli 35 . Several bacterial proteins have demonstrated membrane tubule formation capabilities in vitro 16,36–40 , but despite the growing number of reports, proteins involved in shaping bacterial membranes into OMV/Es have yet to be identified.…”

Section: Introductionmentioning

confidence: 99%

A Prokaryotic Membrane Sculpting BAR Domain Protein

Phillips

Zacharoff

Hampton

et al. 2020

Preprint

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Paragraph1 Bin/Amphiphysin/RVS (BAR) domain proteins belong to a ubiquitous superfamily 2 of coiled-coil proteins that influence membrane curvature in eukaryotes and are 3 associated with vesicle biogenesis, vesicle-mediated protein trafficking, and intracellular 4 signaling 1-6 . BAR domain proteins have not been identified in bacteria, despite certain 5 organisms displaying an array of membrane curvature phenotypes [7][8][9][10][11][12][13][14][15][16] . Here we identify 6 a prokaryotic BAR domain protein, BdpA, from Shewanella oneidensis MR-1, an iron 7 reducing bacterium known to produce redox active membrane vesicles and micrometer-8 scale membrane extensions. BdpA is required for uniform size distribution of outer 9 membrane vesicles and is responsible for scaffolding outer membrane extensions 10 (OMEs) into membrane structures with consistent diameter and curvature. While a strain 11 lacking BdpA produces OMEs, cryogenic transmission electron microscopy reveals more 12 lobed, disordered OMEs rather than the membrane tubes produced by the wild type 13 strain. Overexpression of BdpA promotes OME formation even during planktonic 14 conditions where S. oneidensis OMEs are less common. Heterologous expression also 15 results in OME production in Marinobacter atlanticus CP1 and Escherichia coli. Based 16 on the ability of BdpA to alter membrane curvature in vivo, we propose that BdpA and its 17 homologs comprise a newly identified class of prokaryotic BAR (P-BAR) domains that will 18 aid in identification of putative P-BAR proteins in other bacterial species.curved surface of the antiparallel coiled-coil protein dimers [17][18][19][20] . Some BAR domain- 24containing proteins, such as the N-BAR protein BIN1, contain amphipathic helical wedges 25 that insert into the outer membrane leaflet and can assist in membrane binding 21 . Other 26BAR domains can be accompanied by a membrane targeting domain, such as PX for 27 phosphoinositide binding 22,23 , in order to direct membrane curvature formation at specific 28 sites, as is the case with sorting nexin BAR proteins 4 . The extent of accumulation of BAR 29 domain proteins at a specific site can influence the degree of the resultant membrane 30 curvature 24 , and tubulation events arise as a consequence of BAR domain 31 multimerization in conjunction with lipid binding 25 . Interactions between BAR domain 32 proteins and membranes resolve membrane tension, promote membrane stability, and 33 aid in localizing cellular processes, such as actin binding, signaling through small 34 GTPases, membrane vesicle scission, and vesicular transport of proteins 1,26,27 . Despite 35 our knowledge of numerous eukaryotic BAR proteins spanning a variety of modes of 36 curvature formation, membrane localizations, and subtypes (N-BAR, F-BAR, and I-BAR), 37 characterization of a functional prokaryotic BAR domain protein has yet to be reported. 38 Bacterial cell membrane curvature can be observed during the formation of outer 39 membrane vesicles (OMV) and outer membrane extensions (OME)....

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A Ubiquitous Platform for Bacterial Nanotube Biogenesis

Cited by 41 publications

References 47 publications

Donor-delivered cell wall hydrolases facilitate nanotube penetration into recipient bacteria

Donor-delivered cell wall hydrolases facilitate nanotube penetration into recipient bacteria

The substrate specificity switch FlhB assembles onto the export gate to regulate type three secretion

A Prokaryotic Membrane Sculpting BAR Domain Protein

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