Single Molecule Analysis of RNA Polymerase Elongation Reveals Uniform Kinetic Behavior

By using single-molecule measurements, we demonstrate that the elongation kinetics of individual Escherichia coli RNA polymerase molecules are remarkably homogeneous. We find no evidence of distinct elongation states among RNA polymerases. Instead, the observed heterogeneity in transcription rates r...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2002-10, Vol.99 (21), p.13538-13543
Main Authors: Adelman, Karen, La Porta, Arthur, Santangelo, Thomas J., Lis, John T., Roberts, Jeffrey W., Wang, Michelle D.
Format: Article
Language:English
Subjects:
Antibodies
Bacteria
Biological Sciences
DNA
DNA-Directed RNA Polymerases - chemistry
DNA-Directed RNA Polymerases - genetics
DNA-Directed RNA Polymerases - metabolism
Escherichia coli - enzymology
Escherichia coli - genetics
Genes
Government regulation
Instantaneous velocity
Kinetics
Molecular biology
Molecules
Nucleotides
Plasmids
Point Mutation
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Ribonucleic acid
RNA
Transcription, Genetic
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Summary:By using single-molecule measurements, we demonstrate that the elongation kinetics of individual Escherichia coli RNA polymerase molecules are remarkably homogeneous. We find no evidence of distinct elongation states among RNA polymerases. Instead, the observed heterogeneity in transcription rates results from statistical variation in the frequency and duration of pausing. When transcribing a gene without strong pause sites, RNA polymerase molecules display transient pauses that are distributed randomly in both time and distance. Transitions between the active elongation mode and the paused state are instantaneous within the resolution of our measurements (< 1 s). This elongation behavior is compared with that of a mutant RNA polymerase that pauses more frequently and elongates more slowly than wild type.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.212358999