The role of DNA base excision repair in filamentation in Escherichia coli K-12 adhered to epithelial HEp-2 cells

Costa, Suelen B.; Campos, Ana Carolina da Cruz; Pereira, Ana Claudia Machado; Mattos‐Guaraldi, Ana Luíza; Júnior, Raphael Hirata; Rosa, Ana Cláudia P.; Asad, L.M.B.O.

doi:10.1007/s10482-011-9649-z

Cited by 4 publications

(5 citation statements)

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“…Moreover, this adherence to glass was accompanied by filamentous bacterial growth in the xthA mutant (BW9091) () and especially in the triple xthA nfo nth mutant (BW535) (). This result is similar to that observed in the adherence to HEp-2 cells where BER mutants (BW9091 and BW535) also revealed higher filamentation (Costa et al , 2012). Nevertheless, the wild-type strain (AB1157) presented rare filaments () and the strain deficient in SOS induction ( lexA Ind − , DM49) did not exhibit bacterial cell filamentation ().…”

Section: Resultssupporting

confidence: 88%

“…We showed previously that adherence to HEp-2 cells is accompanied by filamentous growth and SOS induction in a wild-type strain (AB1157), and especially in BER mutants (BW9091 and BW535) (Costa et al , 2012). Therefore, we wondered whether the attachment to glass could lead to DNA damage in the same BER mutants, resulting in a filamentation response.…”

Section: Resultsmentioning

confidence: 95%

“…We showed previously that induction of the SOS response occurs when E. coli K-12 strains interact with epithelial cells (Costa et al , 2012). In this study, we verified that adherence to abiotic surfaces is accompanied by filamentous growth, except in lexA Ind − (DM49) and nfi (1506) mutant strains that did not induce SOS.…”

Section: Discussionmentioning

confidence: 98%

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Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions

et al. 2014

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During the colonization of surfaces, Escherichia coli bacteria often encounter DNA-damaging agents and these agents can induce several defence mechanisms. Base excision repair (BER) is dedicated to the repair of oxidative DNA damage caused by reactive oxygen species (ROS) generated by chemical and physical agents or by metabolism. In this work, we have evaluated whether the interaction with an abiotic surface by mutants derived from E. coli K-12 deficient in some enzymes that are part of BER causes DNA damage and associated filamentation. Moreover, we studied the role of endonuclease V (nfi gene; 1506 mutant strain) in biofilm formation. Endonuclease V is an enzyme that is involved in DNA repair of nitrosative lesions. We verified that endonuclease V is involved in biofilm formation. Our results showed more filamentation in the xthA mutant (BW9091) and triple xthA nfo nth mutant (BW535) than in the wild-type strain (AB1157). By contrast, the mutant nfi did not present filamentation in biofilm, although its wild-type strain (1466) showed rare filaments in biofilm. The filamentation of bacterial cells attaching to a surface was a consequence of SOS induction measured by the SOS chromotest. However, biofilm formation depended on the ability of the bacteria to induce the SOS response since the mutant lexA Ind " did not induce the SOS response and did not form any biofilm. Oxygen tension was an important factor for the interaction of the BER mutants, since these mutants exhibited decreased quantitative adherence under anaerobic conditions. However, our results showed that the presence or absence of oxygen did not affect the viability of BW9091 and BW535 strains. The nfi mutant and its wild-type did not exhibit decreased biofilm formation under anaerobic conditions. Scanning electron microscopy was also performed on the E. coli K-12 strains that had adhered to the glass, and we observed the presence of a structure similar to an extracellular matrix that depended on the oxygen tension. In conclusion, it was proven that bacterial interaction with abiotic surfaces can lead to SOS induction and associated filamentation. Moreover, we verified that endonuclease V is involved in biofilm formation.

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Section: Resultssupporting

confidence: 88%

Section: Resultsmentioning

confidence: 95%

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Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions

et al. 2014

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“…These results are in accordance with the hypothesis that low-intensity laser effects could depend on the genetic characteristics of cells [8,18], in particular those related to DNA repair systems [19,29,30,33]. In fact, it has been reported that the filamentation phenotype was increased for BW9091 and BW535 (exonuclease III, endonuclease III, endonuclease triple mutant) and inhibited by complementation of these mutants with a wild type exonuclease III gene in an assay of cell adherence [45].…”

Section: Discussionsupporting

confidence: 90%

The effects of a low-intensity red laser on bacterial growth, filamentation and plasmid DNA

et al. 2013

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Exposure of nonphotosynthesizing microorganisms to light could increase cell division in cultures, a phenomenon denominated as biostimulation. However, data concerning the importance of the genetic characteristics of cells on this effect are as yet scarce. The aim of this work was to evaluate the effects of a low-intensity red laser on the growth, filamentation and plasmids in Escherichia coli cells proficient and deficient in DNA repair. E. coli cultures were exposed to a laser (658 nm, 10 mW, 1 and 8 J cm−2) to study bacterial growth and filamentation. Also, bacterial cultures hosting pBSK plasmids were exposed to the laser to study DNA topological forms from the electrophoretic profile in agarose gels. Data indicate the low-intensity red laser: (i) had no effect on the growth of E. coli wild type and exonuclease III deficient cells; (ii) induced bacterial filamentation, (iii) led to no alteration in the electrophoretic profile of plasmids from exonuclease III deficient cells, but plasmids from wild type cells were altered. A low-intensity red laser at the low fluences used in phototherapy has no effect on growth, but induces filamentation and alters the topological forms of plasmid DNA in E. coli cultures depending on the DNA repair mechanisms.

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“…3; Additional file 1: Figure S1). Elongated bacterial shape has been observed in stressed conditions such as oxidative stress, nutrient limitation, DNA damages, antibiotics exposure, or extensive recombinant protein expression [29–31] altering DNA replication and cell division. The plausible reason for this might be that the E. coli might have recognized stress and exhibited elongated cell morphology only within a narrow range of stress conditions, i.e., when the growth rate during the chemostat was D = 0.1 h −1 , corresponding to early stationary phase, while the SEM observation and flow cytometry analysis were performed using mid log phase cells in batch cultures.…”

Section: Discussionmentioning

confidence: 99%

An evolutionary optimization of a rhodopsin-based phototrophic metabolism in Escherichia coli

Kim¹,

Kim²,

Park³

et al. 2017

Microb Cell Fact

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BackgroundThe expression of the Gloeobacter rhodopsin (GR) in a chemotrophic Escherichia coli enables the light-driven phototrophic energy generation. Adaptive laboratory evolution has been used for acquiring desired phenotype of microbial cells and for the elucidation of basic mechanism of molecular evolution. To develop an optimized strain for the artificially acquired phototrophic metabolism, an ancestral E. coli expressing GR was adaptively evolved in a chemostat reactor with constant illumination and limited glucose conditions. This study was emphasized at an unexpected genomic mutation contributed to the improvement of microbial performance.ResultsDuring the chemostat culture, increase of cell size was observed, which were distinguished from that of the typical rod-shaped ancestral cells. A descendant ET5 strain was randomly isolated from the chemostat culture at 88-days. The phototrophic growth and the light-induced proton pumping of the ET5 strain were twofold and eightfold greater, respectively, than those of the ancestral E. coli strain. Single point mutation of C1082A at dgcQ gene (encoding diguanylate cyclase, also known as the yedQ gene) in the chromosome of ET5 strain was identified from whole genome sequencing analysis. An ancestral E. coli complemented with the same dgcQ mutation from the ET5 was repeated the subsequently enhancements of light-driven phototrophic growth and proton pumping. Intracellular c-di-GMP, the product of the diguanylate cyclase (dgcQ), of the descendant ET5 strain was suddenly increased while that of the ancestral strain was negligible.ConclusionsNewly acquired phototrophic metabolism of E. coli was further improved via adaptive laboratory evolution by the rise of a point mutation on a transmembrane cell signaling protein followed by increase of signal molecule that eventually led an increase proton pumping and phototrophic growth.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0725-6) contains supplementary material, which is available to authorized users.

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The role of DNA base excision repair in filamentation in Escherichia coli K-12 adhered to epithelial HEp-2 cells

Cited by 4 publications

References 27 publications

Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions

Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions

The effects of a low-intensity red laser on bacterial growth, filamentation and plasmid DNA

An evolutionary optimization of a rhodopsin-based phototrophic metabolism in Escherichia coli

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