Lon Protease Has Multifaceted Biological Functions in
            <i>Acinetobacter baumannii</i>

Ching, Carly; Yang, B. Y.; Onwubueke, Chineme; Lazinski, David W.; Camilli, Andrew; Godoy, Veronica G.

doi:10.1128/jb.00536-18

Cited by 25 publications

(25 citation statements)

References 65 publications

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“…1D). We have previously observed partial complementation of colony biofilms phenotypes with plasmid borne ectopic gene copies, possibly due to the sensitivity of the process to gene dosage (Ching et al, 2018). The data obtained from these experiments suggest that the ∆recA strain forms robust biofilms.…”

Section: Loss Of Reca Promotes Biofilms In a Baumanniimentioning

confidence: 75%

“…To test this and monitor both RecA and csuAB expression we constructed a plasmid borne transcriptional reporter of Csu genes as the one we used previously (Fig. 4), but used instead mCherry, PcsuAB-mCherry , and we introduced it into the A. baumannii strain already containing a chromosomal PrecA-gfp reporter (Ching et al, 2018). 48 hr pellicle biofilms of the double reporter strain were treated with 0.5X MIC of ciprofloxacin (Cip), which is known to induce the DDR but does not kill cells (MacGuire et al, 2014), for an additional 48 hrs.…”

Section: Reca Levels and Csuab Expression Are Inversely Correlated Inmentioning

confidence: 99%

“…The well content was thoroughly mixed before taking the samples, diluted with PBS, and plated on LB agar. After CFU sampling, the well content was aspirated, washed 3 times with PBS, and stained with Crystal Violet as before (Ching et al, 2018). For colony biofilms, 3 µL of a saturated bacterial culture were spotted on to YT agar plates containing 200µg/mL Congo red and 100µg/mL Coomassie brilliant blue (Congo red plates) (Ching et al, 2018) and incubated for 48 hrs., early colony biofilms, to 96 hrs.…”

Section: Pellicle and Colony Biofilm Formationmentioning

confidence: 99%

“…Crystal Violet staining was then carried out as before (Chen et al, 2015). LIVE/DEAD staining was performed on resuspended surface-attached cells, and microscopy performed as before (Ching et al, 2018). The LIVE/DEAD stained surface attached cells of the WT, ∆recA, and recA ++ after Gm treatment (10X MIC) were mechanically dislodged.…”

Section: Mic and Biofilm Eradication Assays And Live/dead Determinatmentioning

confidence: 99%

“…(A) Csu pili are highly expressed in biofilm cells. Confocal microscope imaging of a double label recA-GFP(Ching et al, 2018) and csu mCherry strain. The thickness of the biofilm in the mid-Z stack is 14µm as shown in the left-hand side of the figure.…”

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confidence: 99%

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RecA levels modulate biofilm development inAcinetobacter baumannii

Ching

Muller

Brychcy

et al. 2019

Preprint

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9Acinetobacter baumannii, a Gram-negative opportunistic pathogen, is a leading cause of hospital-10 acquired infections. A. baumannii is difficult to eradicate from hospitals due to its propensity to 11 quickly gain antibiotic resistances and ability to robustly survive on dry surfaces. These strategies are 12 largely mediated by mutagenesis and biofilm development, respectively. Mutagenesis is partly 13 governed by the DNA damage response (DDR). Biofilms are multicellular communities, often 14 surface-attached, that are more difficult to eradicate than free-living planktonic cells. There is 15 increasing evidence that the DDR and biofilm development are linked processes. Here, we show that 16 upon DNA damage, the relative intracellular concentration of RecA, the key DDR protein, is lower 17 than those of Escherichia coli. Notably, we report that RecA negatively influences biofilm 18 development. Cells lacking RecA (∆recA), that are unable to upregulate the DDR, have increased 19 surface attachment and sugar content within the biofilm matrix. We further show that in A. 20 baumannii, a modest increase in RecA concentrations, akin to DDR induction, decreases surface-21 attachment. Importantly, biofilms formed by ∆ recA cells are more difficult to eradicate with 22 2 antibiotic treatment. The evidence suggests that the A. baumannii DDR influences survival 23 independent from mutagenesis. It also demonstrates the importance of understanding fundamental 24 biology to better appreciate the relationships between different bacterial survival strategies. 25 Introduction 26Acinetobacter baumannii is an emerging Gram-negative opportunistic pathogen and one of the 27 ESKAPE pathogens, a group of bacteria responsible for the majority of hospital-acquired infections 28 1 . Interest in these bacteria has stemmed from A. baumannii outbreaks in hospitals worldwide that are 29 difficult to eradicate, due to increased multi-drug resistance (MDR) 2 and resistance to desiccation 3 . 30A. baumannii is very dangerous to immunocompromised individuals, causing different illnesses, 31including pneumonia, septicemia, and wound infections 4 . 32One response pathway that underlies MDR is the DNA damage response (DDR). When gene 33 products involved in antibiotic binding or processing are mutated, resistance is acquired 5 . 34Mutagenesis can result from induction of error-prone DNA polymerase genes which are part of the 35 DDR regulon, a strategy used by different bacteria 6,7 . In Escherichia coli and many other bacteria, 36 the cells' main recombinase, RecA, and the global transcriptional repressor, LexA, manage the DDR, 37 also known as the SOS response 8 . In A. baumannii, however, there is no known LexA homologue, 38 making the A. baumannii DDR circuitry unique 9,10 . We have shown that in A. baumannii expression 39 of multiple error-prone polymerases and clinically-relevant antibiotic resistance acquisition are 40 dependent on RecA 11 . 41A. baumannii is notorious for its ability to robustly survive on surfaces. Under dry conditions, A....

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Section: Loss Of Reca Promotes Biofilms In a Baumanniimentioning

confidence: 75%

Section: Reca Levels and Csuab Expression Are Inversely Correlated Inmentioning

confidence: 99%

Section: Pellicle and Colony Biofilm Formationmentioning

confidence: 99%

Section: Mic and Biofilm Eradication Assays And Live/dead Determinatmentioning

confidence: 99%

mentioning

confidence: 99%

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RecA levels modulate biofilm development inAcinetobacter baumannii

Ching

Muller

Brychcy

et al. 2019

Preprint

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Impact of Bacillus licheniformis from yaks following antibiotic therapy in mouse model

Zeng,

Gong,

Quan

et al. 2024

Appl Microbiol Biotechnol

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One Extra Copy of lon Gene Causes a Dramatic Increase in Actinorhodin Production by Streptomyces coelicolor A3(2)

2019

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ATP-dependent Lon protease plays important roles in different physiological processes, including cellular differentiation of the bacteria and is a part of an important stress response regulon (HspR/HAIR). In Streptomyces, biosynthesis of secondary metabolites starts with cellular differentiation and stress is one of the factor that affect metabolite production. To clarify the effect of Lon protease on secondary metabolite production, we constructed a recombinant strain of Streptomyces coelicolor A3( 2) that has one extra copy of lon gene with its own promoter and transcriptional terminator in its genome. Expression of lon gene in the recombinant strain was determined by quantitative real time (RT-qPCR). Actinorhodin and undecylprodigiosin production of the recombinant cell was measured in liquid R2YE and it was found to produce about 34 times more actinorhodin and 9 times more undecylprodigiosin than the wild-type at 168 h of growth. Development of stable Streptomyces strains capable of producing high amounts of secondary metabolites is valuable for biotechnology industry. One extra copy of lon gene is enough to boost antibiotic production by S. coelicolor A3(2) and this change do not cause any metabolic burden in the cell.

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Lon Protease Has Multifaceted Biological Functions in Acinetobacter baumannii

Cited by 25 publications

References 65 publications

RecA levels modulate biofilm development inAcinetobacter baumannii

RecA levels modulate biofilm development inAcinetobacter baumannii

Impact of Bacillus licheniformis from yaks following antibiotic therapy in mouse model

One Extra Copy of lon Gene Causes a Dramatic Increase in Actinorhodin Production by Streptomyces coelicolor A3(2)

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