Transformation of Bacillus megaterium by electroporation

Moro, Alejandro; Sánchez, Julio César; Serguera, Ché

doi:10.1007/bf00152449

Cited by 4 publications

(3 citation statements)

References 7 publications

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“…All general methods were carried out as described previously 42 . Transformation into Bm OS2 cells was conducted as described 43 . In brief, Bm OS2 cells were grown up to 1.0 OD 600 , washed with electroporation buffer (25% PEG 6000 and 0.1 M sorbitol) and resuspended in 1 ml of electroporation buffer.…”

Section: Methodsmentioning

confidence: 99%

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: Methodsmentioning

confidence: 99%

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

2020

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“…Transformation into Bm (OS2) cells was carried out as previously described ( Moro et al., 1995 ). Bm cells were grown up to 1.0 OD 600 (10 ml).…”

Section: Methodsmentioning

confidence: 99%

A Ubiquitous Platform for Bacterial Nanotube Biogenesis

et al. 2019

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Summary We have previously described the existence of membranous nanotubes, bridging adjacent bacteria, facilitating intercellular trafficking of nutrients, cytoplasmic proteins, and even plasmids, yet components enabling their biogenesis remain elusive. Here we reveal the identity of a molecular apparatus providing a platform for nanotube biogenesis. Using Bacillus subtilis ( Bs ), we demonstrate that conserved components of the flagellar export apparatus (FliO, FliP, FliQ, FliR, FlhB, and FlhA), designated CORE, dually serve for flagellum and nanotube assembly. Mutants lacking CORE genes, but not other flagellar components, are deficient in both nanotube production and the associated intercellular molecular trafficking. In accord, CORE components are located at sites of nanotube emergence. Deleting CORE s of distinct species established that CORE-mediated nanotube formation is widespread. Furthermore, exogenous CORE s from diverse species could restore nanotube generation and functionality in Bs lacking endogenous CORE . Our results demonstrate that the CORE-derived nanotube is a ubiquitous organelle that facilitates intercellular molecular trade across the bacterial kingdom.

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“…All the experiments were carried out at 37 °C. Transformation into Bm OS2 cells was carried out as previously described 37 . In brief, Bm OS2 cells were grown up to 1.0 OD 600 (10 ml).…”

Section: Methodsmentioning

confidence: 99%

Interspecies nutrient extraction and toxin delivery between bacteria

et al. 2017

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Bacteria have developed various mechanisms by which they sense, interact, and kill other bacteria, in an attempt to outcompete one another and survive. Here we show that Bacillus subtilis can kill and prey on Bacillus megaterium. We find that Bacillus subtilis rapidly inhibits Bacillus megaterium growth by delivering the tRNase toxin WapA. Furthermore, utilizing the methionine analogue L-azidohomoalanine as a nutrient reporter, we provide evidence of nutrient extraction from Bacillus megaterium by Bacillus subtilis. Toxin delivery and nutrient extraction occur in a contact-dependent manner, and both activities are abolished in the absence of the phosphodiestrase YmdB, shown previously to mediate intercellular nanotube formation. Furthermore, we detect the localization of WapA molecules to nanotubes. Thus, we propose that Bacillus subtilis utilizes the same nanotube apparatus in a bidirectional manner, delivering toxin and acquiring beneficial cargo, thereby maximally exploiting potential niche resources.

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Transformation of Bacillus megaterium by electroporation

Cited by 4 publications

References 7 publications

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

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

A Ubiquitous Platform for Bacterial Nanotube Biogenesis

Interspecies nutrient extraction and toxin delivery between bacteria

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