scholarly journals Phenotypic Heterogeneity in Sugar Utilization by E. coli Is Generated by Stochastic Dispersal of the General PTS Protein EI from Polar Clusters

2018 ◽  
Vol 8 ◽  
Author(s):  
Sutharsan Govindarajan ◽  
Nitsan Albocher ◽  
Tamar Szoke ◽  
Anat Nussbaum-Shochat ◽  
Orna Amster-Choder
2009 ◽  
Vol 76 (5) ◽  
pp. 1524-1532 ◽  
Author(s):  
Tasha A. Desai ◽  
Christopher V. Rao

ABSTRACT Bacteria such as Escherichia coli will often consume one sugar at a time when fed multiple sugars, in a process known as carbon catabolite repression. The classic example involves glucose and lactose, where E. coli will first consume glucose, and only when it has consumed all of the glucose will it begin to consume lactose. In addition to that of lactose, glucose also represses the consumption of many other sugars, including arabinose and xylose. In this work, we characterized a second hierarchy in E. coli, that between arabinose and xylose. We show that, when grown in a mixture of the two pentoses, E. coli will consume arabinose before it consumes xylose. Consistent with a mechanism involving catabolite repression, the expression of the xylose metabolic genes is repressed in the presence of arabinose. We found that this repression is AraC dependent and involves a mechanism where arabinose-bound AraC binds to the xylose promoters and represses gene expression. Collectively, these results demonstrate that sugar utilization in E. coli involves multiple layers of regulation, where cells will consume first glucose, then arabinose, and finally xylose. These results may be pertinent in the metabolic engineering of E. coli strains capable of producing chemical and biofuels from mixtures of hexose and pentose sugars derived from plant biomass.


Microbiology ◽  
2021 ◽  
Vol 90 (2) ◽  
pp. 237-246
Author(s):  
Z. A. Mirani ◽  
Sh. Urooj ◽  
A. Ullah ◽  
M. N. Khan ◽  
N. Rauf ◽  
...  

2020 ◽  
Author(s):  
Stephen F. Fitzgerald ◽  
Nadejda Lupolova ◽  
Sharif Shaaban ◽  
Timothy J. Dallman ◽  
David Greig ◽  
...  

AbstractThe human zoonotic pathogen Escherichia coli O157 is defined by its extensive prophage repertoire including those that encode Shiga toxin, the factor responsible for inducing life-threatening pathology in humans. As well as introducing genes that can contribute to the virulence of a strain, prophage can enable the generation of large-chromosomal rearrangements (LCRs) by homologous recombination. This work examines the types and frequencies of LCRs across the major lineages of the O157 serogroup and defines the phenotypic consequences of specific structural variants. We demonstrate that LCRs are a major source of genomic variation across all lineages of E. coli O157 and by using both optical mapping and ONT long-read sequencing demonstrate that LCRs are generated in laboratory cultures started from a single colony and particular variants are selected during animal colonisation. LCRs are biased towards the terminus region of the genome and are bounded by specific prophages that share large regions of sequence homology associated with the recombinational activity. RNA transcriptional profiling and phenotyping of specific structural variants indicated that important virulence phenotypes such as Shiga toxin production, type 3 secretion and motility are affected by LCRs. In summary, E. coli O157 has acquired multiple prophage regions over time that act as genome engineers to continually produce structural variants of the genome. This structural variation is a form of epigenetic regulation that generates sub-population phenotypic heterogeneity with important implications for bacterial adaptation and survival.Author SummaryEscherichia coli has an ‘open genome’ and has acquired genetic information over evolutionary time, often in the form of bacteriophages that integrate into the bacterial genome (prophages). E. coli O157 is a clonal serogroup that is found primarily in ruminants such as cattle but can cause life-threatening infections in humans. E. coli O157 isolates contain multiple prophages including those that encode Shiga-like toxins which are responsible for the more serious disease associated with human infections. We show in this study that many of these prophages exhibit large regions of sequence similarity that allow rearrangements to occur in the genome generating structural variants. These occur routinely during bacterial culture in the laboratory and the variants are detected during animal colonization. The variants generated can give the bacteria altered phenotypes, such as increased motility or toxin production which can be selected in specific environments and therefore represent a highly dynamic mechanism to generate variation in bacterial populations without a change in overall gene content.


2018 ◽  
Vol 4 ◽  
pp. 39-42
Author(s):  
Muna Tamang ◽  
Pujan K.C. ◽  
Punya Kumari Koju ◽  
Puspa Lachhimasyu ◽  
Dinesh Dhakal ◽  
...  

Objectives: The study was done with an aim to determine the gain and loss of functions among the actinomycetes mutants induced by sodium azide.Methods: The study was carried out in the laboratory of the Sainik Awasiya Mahavidhayala, Bhaktapur, Nepal from 2016 December to 2017 March. A total of 30 soil samples were collected from Tokha, Bhaktapur area and Godawari area. Actinomycetes were isolated from the soil sample using pour plate technique on selective media; starch casein agar. The isolates were identified by using standard microbiological methods and each isolate was exposed to different concentration of sodium azide to generate mutants. The wild type and mutants were compared in morphology, biochemical reactions and antibiotic susceptibility to test organism to determine the gain and loss of functions.Results: Among 30 samples processed, 20(67%) actinomycetes were isolated, in which 6 (20%) were identified as the Streptomyces spp. A total of 28 mutants were isolated from 6 wild types by exposed at 10ppm, 20ppm, 40ppm, 50ppm, 100ppm concentration of sodium azide. Out of 28 mutants formed, only 10 mutants from sample showed same pigmentation as its wild type while other 18 mutants showed change in their pigmentation. In sugar utilization test, 8 different sugars for 28 mutants each, 56 cases showed Gain of Function (GOF), similarly 44 cases showed Loss of function (LOF). Antibiosis remained unaffected against Pseudomonas i.e. no GOF or LOF was seen. Only 2 cases of LOF against Staphylococcus aureus were seen while there were no cases of LOF in other pathogens. 3 cases of GOF against E. coli, 4 against S Typhi and 4 against S. aureus were observed.Conclusion: The potential of mutant actinomycetes has been realized, and hence opens exciting avenues in the field of biotechnology and biomedical research.


mBio ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. e02938-20
Author(s):  
Manon Barthe ◽  
Josué Tchouanti ◽  
Pedro Henrique Gomes ◽  
Carine Bideaux ◽  
Delphine Lestrade ◽  
...  

ABSTRACTThe glucose-xylose metabolic transition is of growing interest as a model to explore cellular adaption since these molecules are the main substrates resulting from the deconstruction of lignocellulosic biomass. Here, we investigated the role of the XylR transcription factor in the length of the lag phases when the bacterium Escherichia coli needs to adapt from glucose- to xylose-based growth. First, a variety of lag times were observed when different strains of E. coli were switched from glucose to xylose. These lag times were shown to be controlled by XylR availability in the cells with no further effect on the growth rate on xylose. XylR titration provoked long lag times demonstrated to result from phenotypic heterogeneity during the switch from glucose to xylose, with a subpopulation unable to resume exponential growth, whereas the other subpopulation grew exponentially on xylose. A stochastic model was then constructed based on the assumption that XylR availability influences the probability of individual cells to switch to xylose growth. The model was used to understand how XylR behaves as a molecular switch determining the bistability set-up. This work shows that the length of lag phases in E. coli is controllable and reinforces the role of stochastic mechanism in cellular adaptation, paving the way for new strategies for the better use of sustainable carbon sources in bioeconomy.IMPORTANCE For decades, it was thought that the lags observed when microorganisms switch from one substrate to another are inherent to the time required to adapt the molecular machinery to the new substrate. Here, the lag duration was found to be the time necessary for a subpopulation of adapted cells to emerge and become the main population. By identifying the molecular mechanism controlling the subpopulation emergence, we were able to extend or reduce the duration of the lags. This work is of special importance since it demonstrates the unexpected complexity of monoclonal populations during growth on mixed substrates and provides novel mechanistic insights with regard to bacterial cellular adaptation.


Author(s):  
D. E. Philpott ◽  
A. Takahashi

Two month, eight month and two year old rats were treated with 10 or 20 mg/kg of E. Coli endotoxin I. P. The eight month old rats proved most resistant to the endotoxin. During fixation the aorta, carotid artery, basil arartery of the brain, coronary vessels of the heart, inner surfaces of the heart chambers, heart and skeletal muscle, lung, liver, kidney, spleen, brain, retina, trachae, intestine, salivary gland, adrenal gland and gingiva were treated with ruthenium red or alcian blue to preserve the mucopolysaccharide (MPS) coating. Five, 8 and 24 hrs of endotoxin treatment produced increasingly marked capillary damage, disappearance of the MPS coating, edema, destruction of endothelial cells and damage to the basement membrane in the liver, kidney and lung.


Author(s):  
James A. Lake

The understanding of ribosome structure has advanced considerably in the last several years. Biochemists have characterized the constituent proteins and rRNA's of ribosomes. Complete sequences have been determined for some ribosomal proteins and specific antibodies have been prepared against all E. coli small subunit proteins. In addition, a number of naturally occuring systems of three dimensional ribosome crystals which are suitable for structural studies have been observed in eukaryotes. Although the crystals are, in general, too small for X-ray diffraction, their size is ideal for electron microscopy.


Author(s):  
Manfred E. Bayer

Bacterial viruses adsorb specifically to receptors on the host cell surface. Although the chemical composition of some of the cell wall receptors for bacteriophages of the T-series has been described and the number of receptor sites has been estimated to be 150 to 300 per E. coli cell, the localization of the sites on the bacterial wall has been unknown.When logarithmically growing cells of E. coli are transferred into a medium containing 20% sucrose, the cells plasmolize: the protoplast shrinks and becomes separated from the somewhat rigid cell wall. When these cells are fixed in 8% Formaldehyde, post-fixed in OsO4/uranyl acetate, embedded in Vestopal W, then cut in an ultramicrotome and observed with the electron microscope, the separation of protoplast and wall becomes clearly visible, (Fig. 1, 2). At a number of locations however, the protoplasmic membrane adheres to the wall even under the considerable pull of the shrinking protoplast. Thus numerous connecting bridges are maintained between protoplast and cell wall. Estimations of the total number of such wall/membrane associations yield a number of about 300 per cell.


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