Cefodizime: effects of sub-inhibitory concentrations on adhesiveness and bacterial morphology of Staphylococcus aureus and Escherichia coli: comparison with cefotaxime and ceftriaxone

Braga, Pier Carlo; Sasso, M. Dal; Maci, Stefania Maria

doi:10.1093/jac/39.1.79

Cited by 27 publications

(22 citation statements)

References 7 publications

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“…During growth in the absence of IPTG, all mutant cells eventually formed bulges before death. Similar membrane bulging is seen in bacteria treated with cell wall-targeting antibiotics (8,10) and is frequently observed in the E. coli mutant BC202 (35), suggesting that DedA family proteins play a pivotal role in envelope maintenance.…”

Section: Vol 192 2010supporting

confidence: 56%

BB0250 of Borrelia burgdorferi Is a Conserved and Essential Inner Membrane Protein Required for Cell Division

Liang

Sikdar³

et al. 2010

J Bacteriol

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The gene bb0250 of Borrelia burgdorferi is a homolog of the dedA family, encoding integral inner membrane proteins that are present in nearly all species of bacteria. To date, no precise function has been attributed to any dedA gene. Unlike many bacterial species, such as Escherichia coli, which has eight dedA genes, B. burgdorferi possesses only one, annotated bb0250, providing a unique opportunity to investigate the functions of the dedA family. Here, we show that bb0250 is able to restore normal growth and cell division to a temperature-sensitive E. coli mutant with simultaneous deletions of two dedA genes, yqjA and yghB, and encodes a protein that localizes to the inner membrane of E. coli. The bb0250 gene could be deleted from B. burgdorferi only after introduction of a promoterless bb0250 under the control of an inducible lac promoter, indicating that it is an essential gene in this organism. Growth of the mutant in the absence of isopropyl-␤-D-thiogalactopyranoside resulted in cell death, preceded by cell division defects characterized by elongated cells and membrane bulges, demonstrating that bb0250 is required for proper cell division and envelope integrity. Finally, we show that BB0250 depletion leads to imbalanced membrane phospholipid composition in borrelia. These results demonstrate a strong conservation of function of the dedA gene family across diverse species of Gram-negative bacteria and a requirement for this protein family for normal membrane lipid composition and cell division.The dedA family is a highly conserved bacterial gene family encoding inner membrane proteins of unknown function (35). There are more than 2,000 homologs currently found in the NCBI protein database (protein BLAST score versus Escherichia coli DedA of Ͻ0.02), and many species of bacteria have multiple homologs. This built-in redundancy has precluded easy genetic analysis. Each of the dedA homologs in E. coli (yqjA, yghB, yabI, yohD, dedA, ydjX, ydjZ, and yqaA) is individually nonessential as the single gene knockouts have been made and are available in the Keio collection (1). Our group has determined that simultaneous deletion of yghB and yqjA from E. coli results in a strain (named BC202; ⌬yghB::Kan r ⌬yqjA::Tet r ) that has abnormal membrane phospholipid composition, does not complete cell division (forming chains of cells), and fails to grow at 42°C (35). YghB and YqjA are proteins of 219 and 220 amino acids, respectively, displaying 61% amino acid identity. The other six E. coli homologs display roughly 25 to 30% amino acid identity with each other and YghB/YqjA.The E. coli mutant BC202 referred to above displays several intriguing phenotypes that reflect important functions for the DedA family. The membrane and cell division defects of BC202 are present at both the permissive and nonpermissive growth temperatures. However, BC202 is not hypersensitive to antibiotics or detergents, likely signifying an intact outer membrane, under permissive growth conditions (35). We have demonstrated that the periplasmic amidases...

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Section: Vol 192 2010supporting

confidence: 56%

BB0250 of Borrelia burgdorferi Is a Conserved and Essential Inner Membrane Protein Required for Cell Division

Liang

Sikdar³

et al. 2010

J Bacteriol

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“…Other variables known to influence results relate to the test organism and incubation conditions. Bacterial factors affecting morphostructural changes include the cell wall structure of the test organism (Gram-positive vs Gram-negative) [50], the species that is used [51,52], the characteristics of the test strain (including its antibiotic susceptibility) [53,54], and the inoculum density [55,56]. Growthphase affects the size, shape, and cell wall thickness of bacteria even in the absence of antibacterial agent, stationary phase cells being smaller [57,58], having a lower axial ratio [57], and a thicker cell wall [59,60] than logarithmically growing cells.…”

Section: Variables Affecting Antibacterial Agent-induced Morphologicamentioning

confidence: 99%

“…Increases in the diameter of cocci can be difficult to interpret if detected by scanning electron microscopy (SEM) only [50,61]. If, however, the enlarged bacteria are examined in cross-section, the increase in size can usually be attributed to inhibition of cell separation ('pseudomulticellular bacteria') [127,158], or osmotic swelling ('spheroplasts' or 'protoplasts') [84].…”

Section: Altered Cell Sizementioning

confidence: 99%

“…This change in bacterial morphology has been described as 'bulging' by some authors [50,103,169] and 'swelling' by others [55,171,172], with the terms 'oval-centred cells' [54], 'sphero-rods' [101], and 'spindle-shaped forms' [57,173] used for rods with mid-cell swelling, and 'bottle-shaped forms' used for rods with polar swelling [57].…”

Section: Localized Swellingmentioning

confidence: 99%

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Morphological and ultrastructural changes in bacterial cells as an indicator of antibacterial mechanism of action

Cushnie

O’Driscoll

Lamb

2016

Cell. Mol. Life Sci.

159

124

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“…The expression of virulence functions such as toxins, adhesins, and biofilm formation in the human pathogen Staphylococcus aureus is affected by exposure to sub-MICs of antibiotics (2,6,7,11,12,14,19). Sub-MICs of certain antibiotics, in particular, compounds whose primary mode of action is DNA damage, are known to enhance mutation rates in bacteria (15).…”

mentioning

confidence: 99%

Effects of Subinhibitory Concentrations of Antibiotics on SOS and DNA Repair Gene Expression in Staphylococcus aureus

Mesak

Miao

Davies

2008

Antimicrob Agents Chemother

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Reporter clones of Staphylococcus aureus with different SOS response-and DNA repair-associated promoterlux gene fusion constructs were constructed to study the effects of sub-MICs of antibiotics on the transcription of the SOS and methyl mismatch repair (MMR) genes. Fluoroquinolones (FQs) upmodulated both the SOS and the MMR genes. The patterns of antibiotic-induced transcriptional modulation were altered in FQresistant mutants.Subinhibitory concentrations (sub-MICs) of antibiotics are known to provoke extensive transcriptional changes in bacteria (28,29). The expression of virulence functions such as toxins, adhesins, and biofilm formation in the human pathogen Staphylococcus aureus is affected by exposure to sub-MICs of antibiotics (2,6,7,11,12,14,19). Sub-MICs of certain antibiotics, in particular, compounds whose primary mode of action is DNA damage, are known to enhance mutation rates in bacteria (15). This is usually the result of transcriptional changes in the genes responsible for DNA repair and preservation of the integrity of the genome, such as the SOS and methyl mismatch repair (MMR) pathways (10,24,25). DNA polymerases of the SOS system lack intrinsic proofreading activity, which leads to mutations when DNA replication bypasses lesions or errors (24). The MMR system maintains the fidelity of DNA replication by postreplicative correction of base mismatches, small insertions, or deletions (8); a strong mutator phenotype is associated with genetic defects in the MMR system (21).We have studied the effects of sub-MICs of antibiotics of different chemical classes and with different modes of action on the principal mediators of the SOS response in S. aureus, lexA and recA (16) (Fig. 1A). We also examined other known or presumptive SOS response genes, umuC, sosA (SAOUHSC_ 01334), dinB, and recF (9), and known or presumptive MMR genes, mutSL (25, 27), mutS2 (SAOUHSC_01099), mutS3 (SAOUHSC_02276), and mutT (SAOUHSC_00429) (see Table SA in the supplemental material). The respective promoter regions (234 to 661 bp) were amplified by PCR and inserted into pAmiLux (L. R. Mesak et al., unpublished data), a promoter cloning vector, at a BamHI site upstream of a modified Photobacterium luminescens luxABCDE (lux) operon encoding luciferase (luxAB) and fatty acid reductase (luxCDE) from pAL2 (4). The constructs were introduced into S. aureus RN4220 (18), and the effects of the antibiotics on transcription in cells grown in NYE medium (26) were monitored by obtaining luminescence measurements. A single colony of S. aureus from NYE agar was resuspended in 200 l water, mixed with 0.7% agar (1:1,000), and poured as an overlay on NYE agar. Paper disks containing selected antibiotics were placed on the overlay, and the culture was incubated at 37°C. After 20 h, luminescence was detected with a luminograph LB980 photon camera (Berthold). Liquid assays were performed at room temperature in a clear-bottom 96-well plate by using starting cultures with an optical density at 595 nm of 0.150. Luminescence was recorded hourly for ...

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Cefodizime: effects of sub-inhibitory concentrations on adhesiveness and bacterial morphology of Staphylococcus aureus and Escherichia coli: comparison with cefotaxime and ceftriaxone

Cited by 27 publications

References 7 publications

BB0250 of Borrelia burgdorferi Is a Conserved and Essential Inner Membrane Protein Required for Cell Division

BB0250 of Borrelia burgdorferi Is a Conserved and Essential Inner Membrane Protein Required for Cell Division

Morphological and ultrastructural changes in bacterial cells as an indicator of antibacterial mechanism of action

Effects of Subinhibitory Concentrations of Antibiotics on SOS and DNA Repair Gene Expression in Staphylococcus aureus

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