Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells.
In Enterococcus faecalis, sex pheromone-mediated transfer of antibiotic resistance plasmids can occur under unfavorable conditions, for example, when inducing pheromone concentrations are low and inhibiting pheromone concentrations are high. To better understand this paradox, we adapted fluorescence...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2017-07-01
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| Series: | PLoS Genetics |
| Online Access: | http://europepmc.org/articles/PMC5515443?pdf=render |
| _version_ | 1811270981949325312 |
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| author | Rebecca J Breuer Arpan Bandyopadhyay Sofie A O'Brien Aaron M T Barnes Ryan C Hunter Wei-Shou Hu Gary M Dunny |
| author_facet | Rebecca J Breuer Arpan Bandyopadhyay Sofie A O'Brien Aaron M T Barnes Ryan C Hunter Wei-Shou Hu Gary M Dunny |
| author_sort | Rebecca J Breuer |
| collection | DOAJ |
| description | In Enterococcus faecalis, sex pheromone-mediated transfer of antibiotic resistance plasmids can occur under unfavorable conditions, for example, when inducing pheromone concentrations are low and inhibiting pheromone concentrations are high. To better understand this paradox, we adapted fluorescence in situ hybridization chain reaction (HCR) methodology for simultaneous quantification of multiple E. faecalis transcripts at the single cell level. We present direct evidence for variability in the minimum period, maximum response level, and duration of response of individual cells to a specific inducing condition. Tracking of induction patterns of single cells temporally using a fluorescent reporter supported HCR findings. It also revealed subpopulations of rapid responders, even under low inducing pheromone concentrations where the overall response of the entire population was slow. The strong, rapid induction of small numbers of cells in cultures exposed to low pheromone concentrations is in agreement with predictions of a stochastic model of the enterococcal pheromone response. The previously documented complex regulatory circuitry controlling the pheromone response likely contributes to stochastic variation in this system. In addition to increasing our basic understanding of the biology of a horizontal gene transfer system regulated by cell-cell signaling, demonstration of the stochastic nature of the pheromone response also impacts any future efforts to develop therapeutic agents targeting the system. Quantitative single cell analysis using HCR also has great potential to elucidate important bacterial regulatory mechanisms not previously amenable to study at the single cell level, and to accelerate the pace of functional genomic studies. |
| first_indexed | 2024-04-12T22:13:06Z |
| format | Article |
| id | doaj.art-76f3355d5f75453fb86d73f42cb8a72f |
| institution | Directory Open Access Journal |
| issn | 1553-7390 1553-7404 |
| language | English |
| last_indexed | 2024-04-12T22:13:06Z |
| publishDate | 2017-07-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj.art-76f3355d5f75453fb86d73f42cb8a72f2022-12-22T03:14:40ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042017-07-01137e100687810.1371/journal.pgen.1006878Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells.Rebecca J BreuerArpan BandyopadhyaySofie A O'BrienAaron M T BarnesRyan C HunterWei-Shou HuGary M DunnyIn Enterococcus faecalis, sex pheromone-mediated transfer of antibiotic resistance plasmids can occur under unfavorable conditions, for example, when inducing pheromone concentrations are low and inhibiting pheromone concentrations are high. To better understand this paradox, we adapted fluorescence in situ hybridization chain reaction (HCR) methodology for simultaneous quantification of multiple E. faecalis transcripts at the single cell level. We present direct evidence for variability in the minimum period, maximum response level, and duration of response of individual cells to a specific inducing condition. Tracking of induction patterns of single cells temporally using a fluorescent reporter supported HCR findings. It also revealed subpopulations of rapid responders, even under low inducing pheromone concentrations where the overall response of the entire population was slow. The strong, rapid induction of small numbers of cells in cultures exposed to low pheromone concentrations is in agreement with predictions of a stochastic model of the enterococcal pheromone response. The previously documented complex regulatory circuitry controlling the pheromone response likely contributes to stochastic variation in this system. In addition to increasing our basic understanding of the biology of a horizontal gene transfer system regulated by cell-cell signaling, demonstration of the stochastic nature of the pheromone response also impacts any future efforts to develop therapeutic agents targeting the system. Quantitative single cell analysis using HCR also has great potential to elucidate important bacterial regulatory mechanisms not previously amenable to study at the single cell level, and to accelerate the pace of functional genomic studies.http://europepmc.org/articles/PMC5515443?pdf=render |
| spellingShingle | Rebecca J Breuer Arpan Bandyopadhyay Sofie A O'Brien Aaron M T Barnes Ryan C Hunter Wei-Shou Hu Gary M Dunny Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. PLoS Genetics |
| title | Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. |
| title_full | Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. |
| title_fullStr | Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. |
| title_full_unstemmed | Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. |
| title_short | Stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells. |
| title_sort | stochasticity in the enterococcal sex pheromone response revealed by quantitative analysis of transcription in single cells |
| url | http://europepmc.org/articles/PMC5515443?pdf=render |
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