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Related Concept Videos

Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
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Complementary DNA01:44

Complementary DNA

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Overview
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Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
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RNA Polymerase II Accessory Proteins02:36

RNA Polymerase II Accessory Proteins

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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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Related Experiment Video

Updated: Feb 3, 2026

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
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Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

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E. coli DNA polymerase I as a reverse transcriptase

M Ricchetti1, H Buc

  • 1Unité de Physicochimie des Macromolécules Biologiques (URA 1149 du CNRS), Institut Pasteur, Paris, France.

The EMBO Journal
|February 1, 1993
PubMed
Summary

Escherichia coli DNA polymerase I can retrotranscribe RNA, though its efficiency is limited by low processivity. This enzyme exhibits high accuracy on RNA templates, similar to its DNA replication fidelity.

Area of Science:

  • Molecular Biology
  • Enzymology
  • Biochemistry

Background:

  • DNA polymerase I from Escherichia coli is a key enzyme in DNA replication and repair.
  • Its ability to interact with and process RNA templates is less understood.
  • Understanding retrotranscription capabilities sheds light on enzyme versatility and error-checking mechanisms.

Purpose of the Study:

  • To investigate the retrotranscription capabilities of Escherichia coli DNA polymerase I on RNA templates.
  • To determine the kinetic parameters and processivity of the enzyme when using RNA as a template.
  • To assess the accuracy and fidelity of DNA polymerase I during RNA retrotranscription.

Main Methods:

  • Primer extension assays were employed under steady-state conditions.

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Increasing cDNA Yields from Single-cell Quantities of mRNA in Standard Laboratory Reverse Transcriptase Reactions using Acoustic Microstreaming
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  • Kinetic constants (equilibrium, rate, Michaelis constants) were determined for enzyme binding and substrate incorporation.
  • The role of divalent cations (Magnesium vs. Manganese) in fidelity was examined.

    • Main Results:

    • Escherichia coli DNA polymerase I effectively retrotranscribes RNA templates, showing similar initial binding characteristics to DNA templates.
    • Enzyme processivity limits the maximal velocity for both DNA and RNA templates.
    • Catalytic efficiency on RNA is lower, indicated by a 100-fold higher Michaelis constant for deoxynucleotide substrates.
    • The enzyme demonstrates high accuracy on RNA, with error detection requiring Manganese substitution for Magnesium.
    • Impaired elongation of mismatched primer termini and a very low forward polymerization rate contribute to fidelity.

    Conclusions:

    • Escherichia coli DNA polymerase I retains its fundamental characteristics, including low processivity and high fidelity, when retrotranscribing RNA.
    • The enzyme's intrinsic accuracy is maintained, suggesting conserved mechanisms for preventing errors during nucleic acid synthesis.
    • These findings highlight the enzyme's adaptability and robust error-correction capabilities across different template types.