Induction of L-form-like cell shape change of Bacillus subtilis under microculture conditions

Shingaki, Ryuji; Kasahara, Yasuhiro; Iwano, Megumi; Kuwano, Masayoshi; Takatsuka, Tetsuharu; Inoue, Tetsuyoshi; Kokeguchi, Susumu; Fukui, Kiichi

doi:10.1099/mic.0.26259-0

Cited by 10 publications

(7 citation statements)

References 38 publications

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“…S9). Such morphological changes in B. subtilis cells upon oxygen depletion have been documented before (29,30) and can now be explained by the importance of the membrane potential for protein localization.…”

Section: Resultsmentioning

confidence: 67%

Membrane potential is important for bacterial cell division

Strahl

Hamoen

2010

Proc. Natl. Acad. Sci. U.S.A.

457

463

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Many cell division-related proteins are located at specific positions in the bacterial cell, and this organized distribution of proteins requires energy. Here, we report that the proton motive force, or more specifically the (trans)membrane potential, is directly involved in protein localization. It emerged that the membrane potential modulates the distribution of several conserved cell division proteins such as MinD, FtsA, and the bacterial cytoskeletal protein MreB. We show for MinD that this is based on the membrane potential stimulated binding of its C-terminal amphipathic helix. This function of the membrane potential has implications for how these morphogenetic proteins work and provide an explanation for the effects observed with certain antimicrobial compounds.

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Section: Resultsmentioning

confidence: 67%

Membrane potential is important for bacterial cell division

Strahl

Hamoen

2010

Proc. Natl. Acad. Sci. U.S.A.

457

463

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“…The typical colonies were fixed by glutaraldehyde before being processed and examined by electron microscopy (EM). Scanning EM and transmission EM were performed with scanning electron microscope (JSM-6380Lv) and transmission electron microscope (JEM-1230), respectively, using procedures as described [7] , [21] .…”

Section: Methodsmentioning

confidence: 99%

Glycerol Uptake Is Important for L-Form Formation and Persistence in Staphylococcus aureus

Han

Shi

et al. 2014

PLoS ONE

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S. aureus is a significant human pathogen and has previously been shown to form cell wall deficient forms or L-forms in vitro and in vivo during infection. Despite many previous studies on S. aureus L-forms, the mechanisms of L-form formation in this organism remain unknown. Here we established the L-form model in S. aureus and constructed a transposon mutant library to identify genes involved in L-form formation. Screening of the library for mutants defective in L-form formation identified glpF involved in glycerol uptake being important for L-form formation in S. aureus. Consistent with this observation, glpF was found to be highly expressed in L-form S. aureus but hardly expressed in normal walled form. In addition, glpF mutant was found to be defective in antibiotic persistence. The defect in L-form formation and antibiotic persistence of the glpF mutant could be complemented by the wild type glpF gene. These findings provide new insight into the mechanisms of L-form formation and persistence in S. aureus and may have implications for development of new drugs targeting persisters for improved treatment.

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“…Photos were processed using SPOT software. Transmission electron microscopy and freeze substitution of L-form colonies was performed as described [40].…”

Section: Methodsmentioning

confidence: 99%

Insights into the Molecular Basis of L-Form Formation and Survival in Escherichia coli

2009

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L-forms have been shown to occur among many species of bacteria and are suspected to be involved in persistent infections. Since their discovery in 1935, numerous studies characterizing L-form morphology, growth, and pathogenic potential have been conducted. However, the molecular mechanisms underlying the formation and survival of L-forms remain unknown. Using unstable L-form colonies of Escherichia coli as a model, we performed genome-wide transcriptome analysis and screened a deletion mutant library to study the molecular mechanisms involved in formation and survival of L-forms. Microarray analysis of L-form versus classical colonies revealed many up-regulated genes of unknown function as well as multiple over-expressed stress pathways shared in common with persister cells and biofilms. Mutant screens identified three groups of mutants which displayed varying degrees of defects in L-form colony formation. Group 1 mutants, which showed the strongest defect in L-form colony formation, belonged to pathways involved in cell envelope stress, DNA repair, iron homeostasis, outer membrane biogenesis, and drug efflux/ABC transporters. Four (Group 1) mutants, rcsB, a positive response regulator of colanic acid capsule synthesis, ruvA, a recombinational junction binding protein, fur, a ferric uptake regulator and smpA a small membrane lipoprotein were selected for complementation. Complementation of the mutants using a high-copy overexpression vector failed, while utilization of a low-copy inducible vector successfully restored L-form formation. This work represents the first systematic genetic evaluation of genes and pathways involved in the formation and survival of unstable L-form bacteria. Our findings provide new insights into the molecular mechanisms underlying L-form formation and survival and have implications for understanding the emergence of antibiotic resistance, bacterial persistence and latent infections and designing novel drugs and vaccines.

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Induction of L-form-like cell shape change of Bacillus subtilis under microculture conditions

Cited by 10 publications

References 38 publications

Membrane potential is important for bacterial cell division

Membrane potential is important for bacterial cell division

Glycerol Uptake Is Important for L-Form Formation and Persistence in Staphylococcus aureus

Insights into the Molecular Basis of L-Form Formation and Survival in Escherichia coli

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