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PMID: 15967986 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Real-time RNA profiling within a single bacterium.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 26 ·2005-06-28 ·Pages 9160-4

Le TT, Harlepp S, Guet CC, Dittmar K, Emonet T, Pan T, Cluzel P

Abstract

Characterizing the dynamics of specific RNA levels requires real-time RNA profiling in a single cell. We show that the combination of a synthetic modular genetic system with fluorescence correlation spectroscopy allows us to directly measure in real time the activity of any specific promoter in prokaryotes. Using a simple inducible gene expression system, we found that induced RNA levels within a single bacterium of Escherichia coli exhibited a pulsating profile in response to a steady input of inducer. The genetic deletion of an efflux pump system, a key determinant of antibiotic resistance, altered the pulsating transcriptional dynamics and caused overexpression of induced RNA. In contrast with population measurements, real-time RNA profiling permits identifying relationships between genotypes and transcriptional dynamics that are accessible only at the level of the single cell.

MeSH Terms
Bacterial Proteins/chemistry Cell Cycle Cell Proliferation Dose-Response Relationship, Drug Drug Resistance, Multiple Escherichia coli/metabolism Escherichia coli Proteins/chemistry Gene Expression Regulation, Bacterial Genes, Reporter Genotype Green Fluorescent Proteins/metabolism Models, Statistical Plasmids/metabolism Promoter Regions, Genetic Protein Binding RNA/chemistry,metabolism RNA, Messenger/metabolism Recombinant Fusion Proteins/chemistry Spectrometry, Fluorescence Spliceosomes/metabolism Time Factors Transcription, Genetic
Chemicals
Bacterial Proteins Escherichia coli Proteins RNA, Messenger Recombinant Fusion Proteins Green Fluorescent Proteins RNA
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Le Thuc T
Institute for Biophysical Dynamics and The James Franck Institute and Department of Biochemistry and Molecular Biology, University of Chicago, 5640 South Ellis Avenue, Chicago, IL 60637, USA.
Harlepp Sébastien
Guet Calin C
Dittmar Kimberly
Emonet Thierry
Pan Tao
Cluzel Philippe
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0503311102
Published
2005-06-28
Epub
2005-00-20
Pages
9160-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1166617
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0503311102 PMC Full Text View journal details
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