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

Selection for active E. coli tRNA(Phe) variants from a randomized library using two proteins

E T Peterson1, J Blank, M Sprinzl

  • 1Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.

The EMBO Journal
|July 1, 1993
PubMed
Summary

In vitro selection identified active Escherichia coli tRNA(Phe) variants, revealing broad sequence diversity recognized by phenylalanyl-tRNA synthetase (FRS). This method also uncovered novel tertiary interactions and variant classes, demonstrating FRS

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

Assessment of Knowledge Levels in a Blended-Learning CPD Programme in Periodontology: A Cross-Sectional Comparative Study.

European journal of dental education : official journal of the Association for Dental Education in Europe·2026
Same author

MasterOnline Periodontology and Implant Therapy-revisited after seven years: A case study of the structures and outcomes in a blended learning CPD.

European journal of dental education : official journal of the Association for Dental Education in Europe·2016
Same author

Silencing vimentin expression decreases pulmonary metastases in a pre-diabetic mouse model of mammary tumor progression.

Oncogene·2016
Same author

Structural Studies of Model RNA Helices with Relevance to Aminoacyl-tRNA Synthetase Specificity and HIV Reverse Transcription.

Journal of biomolecular structure & dynamics·2012
Same author

Residency differentials in Mormon fertility.

Population studies·2011
Same author

[Implementing in-patient smoking cessation interventions. Basic characteristics of smoking patients in a lung health department].

Pneumologie (Stuttgart, Germany)·2011

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Transfer RNA (tRNA) molecules are crucial for protein synthesis, acting as adaptors between mRNA codons and amino acids.
  • Phenylalanyl-tRNA synthetase (FRS) is an enzyme responsible for attaching phenylalanine to its cognate tRNA(Phe).
  • Understanding tRNA-enzyme interactions is key to deciphering translational fidelity and regulation.

Purpose of the Study:

  • To isolate and characterize active Escherichia coli tRNA(Phe) variants with altered recognition properties.
  • To investigate the role of specific nucleotide residues and tertiary interactions in FRS recognition.
  • To explore the sequence diversity tolerated by FRS and identify novel tRNA structures.

Main Methods:

  • In vitro selection using iterative rounds of binding to Escherichia coli phenylalanyl-tRNA synthetase.

Related Experiment Videos

  • Aminoacylation assays to confirm functional tRNA activity.
  • Selection for high affinity to elongation factor-Tu.
  • Randomization of specific nucleotide positions within tRNA(Phe) libraries.

    • Main Results:

    • Confirmed the importance of previously identified residues (U20, G34, A35, A36, U59) for FRS recognition.
    • Identified active tRNA(Phe) variants that do not require canonical tertiary interactions (G10-C25-U45, A26-G44).
    • Discovered novel nucleotide combinations at conserved tertiary interaction sites, suggesting new interaction possibilities.
    • Isolated active tRNA variants with deletions, indicating FRS tolerance for structural variations.
    • Identified a second class of variants due to an enzyme preparation impurity, highlighting method sensitivity.

    Conclusions:

    • Escherichia coli phenylalanyl-tRNA synthetase exhibits a surprisingly broad substrate sequence diversity.
    • In vitro selection is a powerful tool for dissecting tRNA-enzyme interactions and discovering novel tRNA functionalities.
    • Canonical tertiary interactions are not strictly essential for all functional tRNA(Phe)-FRS complexes.