Failsafe Mechanisms Couple Division and DNA Replication in Bacteria

Arjes, Heidi A.; Kriel, Allison; Sorto, Nohemy A.; Shaw, Jared T.; Wang, Jue D.; Levin, Petra Anne

doi:10.1016/j.cub.2014.07.055

Cited by 46 publications

(33 citation statements)

References 29 publications

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“…Some distant connections were intuitive, such as DNA replication ( holB ) and folate biosynthesis ( folC and the sul-folB-folK operon), likely reflecting the folate requirement for dTTP production (Hardy et al, 1987). Other connections were unexpected, such as those between peptidoglycan (PG) biosynthesis/cell division and DNA replication/modification (e.g., ftsL and dnaX , murC and gyrB , ddl-murF and ydiO ); these connections may be involved in failsafe mechanisms that link division and DNA replication (Arjes et al, 2014). …”

Section: Resultsmentioning

confidence: 99%

A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria

Peters

Colavin

Shi

et al. 2016

Cell

614

681

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Summary Essential gene functions underpin the core reactions required for cell viability, but their contributions and relationships are poorly studied in vivo. Using CRISPR interference, we created knockdowns of every essential gene in Bacillus subtilis and probed their phenotypes. Our high-confidence essential gene network, established using chemical genomics, showed extensive interconnections among distantly related processes and identified modes of action for uncharacterized antibiotics. Importantly, mild knockdown of essential gene functions significantly reduced stationary phase survival without affecting maximal growth rate, suggesting that essential protein levels are set to maximize outgrowth from stationary phase. Finally, high-throughput microscopy indicated that cell morphology is relatively insensitive to mild knockdown but profoundly affected by depletion of gene function, revealing intimate connections between cell growth and shape. Our results provide a framework for systematic investigation of essential gene functions in vivo that is broadly applicable to diverse microorganisms and amenable to comparative analysis.

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Section: Resultsmentioning

confidence: 99%

A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria

Peters

Colavin

Shi

et al. 2016

Cell

614

681

View full text Add to dashboard Cite

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“…Second, most genetic and cytological data places initial FtsZ ring assembly steps very early in the replication period far in advance of termination (e.g., Addinall and Lutkenhaus, 1996 ; Yu et al, 1998 ; Harry et al, 1999 ). Inhibiting DNA replication prior to or soon after the initiation step imparts a strong cell division block independent of nucleoid occlusion, but once established, inhibition of replication elongation through drug treatment or temperature sensitive replisome mutant does not itself inhibit FtsZ ring assembly (Harry et al, 1999 ; Regamey et al, 2000 ; Arjes et al, 2014 ; Morigen et al, 2014 ). In such a cell, Z-rings form off-center in a nucleoid occlusion-dependent process, generating anucleate cells (Mulder and Woldringh, 1989 ).…”

Section: Introductionmentioning

confidence: 99%

MioC and GidA proteins promote cell division in E. coli

et al. 2015

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The well-conserved genes surrounding the E. coli replication origin, mioC and gidA, do not normally affect chromosome replication and have little known function. We report that mioC and gidA mutants exhibit a moderate cell division inhibition phenotype. Cell elongation is exacerbated by a fis deletion, likely owing to delayed replication and subsequent cell cycle stress. Measurements of replication initiation frequency and origin segregation indicate that mioC and gidA do not inhibit cell division through any effect on oriC function. Division inhibition is also independent of the two known replication/cell division checkpoints, SOS and nucleoid occlusion. Complementation analysis indicates that mioC and gidA affect cell division in trans, indicating their effect is at the protein level. Transcriptome analysis by RNA sequencing showed that expression of a cell division septum component, YmgF, is significantly altered in mioC and gidA mutants. Our data reveal new roles for the gene products of gidA and mioC in the division apparatus, and we propose that their expression, cyclically regulated by chromatin remodeling at oriC, is part of a cell cycle regulatory program coordinating replication initiation and cell division.

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“…2C, S1C). This uncoupling between OD and viability has been observed previously in cell-cycle mutants where cells remain intact and metabolically active but cannot divide and form colonies [51], and may be more prevalent than is currently appreciated, motivating future studies that rely on OD to perform assays to verify culture viability.…”

Section: Discussionmentioning

confidence: 85%

Biosurfactant production maintains viability in anoxic conditions by depolarizing the membrane inBacillus subtilis

Arjes

Dunn

et al. 2019

Preprint

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20membrane potential, membrane energetics, LytC 21 35 depolarizes the B. subtilis membrane, and that other known membrane-potential 36 disruptors restore viability to 168. These findings highlight the importance of surfactin for 37 survival during oxygen-depleted conditions and demonstrate that antimicrobials normally 38 considered harmful can instead benefit cells in stressful conditions when the terminal 39 electron acceptor in respiration is limiting. 40 following a rain [12]. Early observations of B. subtilis culture lysis upon a shift to anoxic 64 environments have yet to be further characterized [13], and it remains a mystery whether 65 B. subtilis has strategies to cope with oxygen limitation. 66 B. subtilis has long been domesticated in the lab, leading to a multitude of genetic 67 tools, strain libraries, and online databases and resources [14][15][16]. The high level of genetic 68 relatedness between biofilm-forming "wild" strains and derivative non-biofilm-forming 69 laboratory strains has been exploited to identify genetic differences that underlie biofilm 70 community behaviors [17]. For instance, the commonly studied laboratory strain 168, 71 which is derived from the biofilm-forming strain 3610, lacks an extrachromosomal plasmid 72 and harbors several point mutations that reduce or abolish social behaviors such as matrix 73 production [18]. Notably, 3610 produces the small molecule surfactin, a powerful 74 surfactant that has previously been implicated in swarming motility [19][20][21]. Surfactin has 75 also been shown to kill fungi [22] and some bacteria in vitro [22][23][24][25][26]. However, fitness 76 benefits of surfactin production in the context of planktonic cultures have yet to be 77 identified, although some surfactants can accelerate oxygen diffusion through the air-water 78 interface [27]. 79 Here, we characterize the interplay between oxygen availability and surfactin 80 production during the growth and death of B. subtilis cultures. We show that oxygen 81 becomes limiting in the culture during the transition to stationary phase and that surfactin 82 secretion improves growth yield in stationary phase by increasing oxygen availability. 83 During a shift to anoxic conditions, we demonstrate that the majority of B. subtilis cells die 84 and lyse due to the activity of the LytC autolysin and surfactin. Finally, we discover that 85 surfactin maintains the viability of the remaining cells by causing membrane 86 depolarization that allows these cells to survive until oxygen is restored. 87 131 hour (Fig. 1D). Since the lysis behavior varied with initial OD, we standardized all further 132 experiments by growing 3610 cells to an OD600 of ~0.9-1.1 before cutting off oxygen 133

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Failsafe Mechanisms Couple Division and DNA Replication in Bacteria

Cited by 46 publications

References 29 publications

A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria

A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria

MioC and GidA proteins promote cell division in E. coli

Biosurfactant production maintains viability in anoxic conditions by depolarizing the membrane inBacillus subtilis

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