Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Function of the ribosomal E-site: a mutagenesis study.

Petr V Sergiev1, Dmitry V Lesnyak, Sergey V Kiparisov

  • 1Department of Chemistry and A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119899, Russia.

Nucleic Acids Research
|October 26, 2005
PubMed
Summary

Mutating Escherichia coli ribosomes destabilized the E-site, leading to increased frameshifting and stop codon readthrough during protein synthesis. This highlights the E-site's crucial role in accurate translation.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Impact of Genetically Induced Metabolic Syndrome on the Outcomes of Photothrombotic Stroke in Leptin-Deficient and Leptin Receptor-Deficient Mice.

Biochemistry. Biokhimiia·2026
Same author

The Spermidine Synthase Gene as a Reporter of Transcription Inhibition in <i>Escherichia coli</i>.

International journal of molecular sciences·2026
Same author

HR-LCMS/MS-Based Dereplication of Plant-Derived Autophagy Inducers Revealed <i>Astragalus dasyanthus</i> as a New Glabrol Producer.

Metabolites·2026
Same author

Novel examples of NMD escape through alternative intronic polyadenylation.

NAR genomics and bioinformatics·2026
Same author

Spirocyclohexane-Chroman-4-one Derivatives as Selectively Cytotoxic Agents in Breast Cancer Models.

ACS medicinal chemistry letters·2026
Same author

PrfH accelerates repair of colicin E3 - damaged translating ribosomes by RtcB2.

RNA biology·2026

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Ribosomes are essential molecular machines responsible for protein synthesis, translating messenger RNA (mRNA) into polypeptide chains.
  • Transfer RNA (tRNA) molecules are critical adaptors, carrying amino acids to the ribosome and interacting with specific binding sites: A-site (aminoacyl-tRNA), P-site (peptidyl-tRNA), and E-site (exit).

Purpose of the Study:

  • To investigate the functional consequences of destabilizing the E-site binding in Escherichia coli ribosomes.
  • To elucidate the role of the E-site in maintaining translational fidelity and efficiency.

Main Methods:

  • A specific mutation (C2394G) in the 23S ribosomal RNA (rRNA) was introduced to destabilize the E-site binding in Escherichia coli.
  • The effects of the mutant rRNA were assessed in vivo through analysis of frameshifting and stop codon readthrough.

Related Experiment Videos

  • In vitro studies examined the ribosomal elongation cycle, tRNA ejection, translocation efficiency, and the interaction with elongation factor G (EF-G).

    • Main Results:

    • Expression of the mutant 23S rRNA resulted in significantly increased frameshifting and stop codon readthrough, indicating reduced translational accuracy.
    • In vitro experiments showed that deacylated tRNA is ejected during or shortly after the translocation step in mutant ribosomes.
    • The mutation impaired the formation of the P/E hybrid site and diminished the stimulation of EF-G GTPase activity by ribosome-bound deacylated tRNA.

    Conclusions:

    • Destabilization of the ribosomal E-site binding compromises translational fidelity, leading to errors like frameshifting and readthrough.
    • The E-site plays a critical role in the efficient ejection of deacylated tRNA during translocation and in regulating elongation factor activity.