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      Associations between cellular levels of ATP and prodigiosin pigment throughout the growth cycle of Serratia marcescens

      1 , 1
      Canadian Journal of Microbiology
      Canadian Science Publishing

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          Abstract

          Serratia marcescens is a prolific producer of the red, membrane-associated pigment prodigiosin. Earlier work has established both a positive role for prodigiosin in ATP production during the population lag phase and a negative role during high-rate, low cell density growth. This study uses the growth rate and growth phase modulation afforded by chemostat culture to extend prodigiosin functional analysis to the high-density and stationary phases. Cellular levels of prodigiosin were positively associated with cellular levels of ATP during high-density growth, and artificial pigment induction during this phase increased cellular ATP levels. Following peak high-density ATP per cell, the early stationary phase enabled significant population growth, while prodigiosin levels remained high and ATP declined. During the late stationary phase, ATP per cell was positively associated with prodigiosin per cell, while both declined during continued growth. These results provide correlational evidence for the multiple effects of prodigiosin pigment on ATP production throughout the growth cycle. Earlier work and the data presented here enable the formulation of a working model for the oscillating relationships between cellular levels of ATP and prodigiosin during batch culture.

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          Persister formation in Staphylococcus aureus is associated with ATP depletion

          Persisters are dormant phenotypic variants of bacterial cells that are tolerant to killing by antibiotics 1 . Persisters are associated with chronic infections and antibiotic treatment failure 1–3 . In Escherichia coli, toxin/antitoxin (TA) modules have been linked to persister formation 4–6 . The mechanism of persister formation in Gram-positive bacteria is unknown. Staphylococcus aureus is a major human pathogen, responsible for a variety of chronic and relapsing infections such as osteomyelitis, endocarditis and infections of implanted devices. Deleting TA modules in S. aureus did not affect the level of persisters. Here we show that S. aureus persisters are produced due to a stochastic entrance into stationary phase accompanied by a drop in intracellular ATP. Cells expressing stationary state markers are present throughout the growth phase, increasing in frequency with cell density. Cell sorting revealed that expression of stationary markers is associated with a 100–1000 fold increase in the likelihood of survival to antibiotic challenge. The ATP level of the cell is predictive of bactericidal antibiotic efficacy and explains bacterial tolerance to antibiotics.
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            Characterization of nucleotide pools as a function of physiological state in Escherichia coli.

            Using a modified method that involves minimal manipulation of cells, we report new information about nucleotide pool sizes and changes throughout the Escherichia coli growth curve. Nucleotide pool sizes are critically dependent on sample manipulation and extraction methods. Centrifugation and even short (2 min) lapses in sample preparation can dramatically affect results. The measured ATP concentration at three different growth rates is at least 3 mM, well above the 0.8 mM needed to saturate the rRNA promoter P1 in vitro. Many of the pools, including ATP, GTP, and UTP, begin to decrease while the cells are still in mid-log growth. After an almost universal drop in nucleotide concentration as the cells transition from logarithmic to stationary phase, there is a "rebound" of certain nucleotides, most notably ATP, after the cells enter stationary phase, followed by a progressive decrease. UTP, in contrast, increases as the cells transition into stationary phase. The higher UTP values might be related to elevated UDP-glucose/galactose, which was found to be at higher concentrations than expected in stationary phase. dTTP is the most abundant deoxynucleoside triphosphate (dNTP) in the cell despite the fact that its precursors, UDP and UTP, are not. All dNTPs decrease through the growth curve but do not have the abrupt drop, as seen with other nucleotides when the cells transition into stationary phase.
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              Genome Evolution and Plasticity of Serratia marcescens, an Important Multidrug-Resistant Nosocomial Pathogen

              Serratia marcescens is an important nosocomial pathogen that can cause an array of infections, most notably of the urinary tract and bloodstream. Naturally, it is found in many environmental niches, and is capable of infecting plants and animals. The emergence and spread of multidrug-resistant strains producing extended-spectrum or metallo beta-lactamases now pose a threat to public health worldwide. Here we report the complete genome sequences of two carefully selected S. marcescens strains, a multidrug-resistant clinical isolate (strain SM39) and an insect isolate (strain Db11). Our comparative analyses reveal the core genome of S. marcescens and define the potential metabolic capacity, virulence, and multidrug resistance of this species. We show a remarkable intraspecies genetic diversity, both at the sequence level and with regards genome flexibility, which may reflect the diversity of niches inhabited by members of this species. A broader analysis with other Serratia species identifies a set of approximately 3,000 genes that characterize the genus. Within this apparent genetic diversity, we identified many genes implicated in the high virulence potential and antibiotic resistance of SM39, including the metallo beta-lactamase and multiple other drug resistance determinants carried on plasmid pSMC1. We further show that pSMC1 is most closely related to plasmids circulating in Pseudomonas species. Our data will provide a valuable basis for future studies on S. marcescens and new insights into the genetic mechanisms that underlie the emergence of pathogens highly resistant to multiple antimicrobial agents.
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                Author and article information

                Journal
                Canadian Journal of Microbiology
                Can. J. Microbiol.
                Canadian Science Publishing
                0008-4166
                1480-3275
                September 2021
                September 2021
                : 67
                : 9
                : 639-650
                Affiliations
                [1 ]Department of Biology, Auburn University at Montgomery, P.O. Box 244023, Montgomery, AL 36124-4023 USA.
                Article
                10.1139/cjm-2020-0619
                f0b77403-9b11-4f5a-8e7d-a668ede92fad
                © 2021

                http://www.nrcresearchpress.com/page/about/CorporateTextAndDataMining

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