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

Construction and processing of transfer RNA precursor models.

C K Surratt1, Z Lesnikowski, A L Schifman

  • 1Department of Chemistry, University of Virginia, Charlottesville 22901.

The Journal of Biological Chemistry
|December 25, 1990
PubMed
Summary
This summary is machine-generated.

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

Mixed infections with distinct cytomegalovirus glycoprotein B genotypes in Polish pregnant women, fetuses, and newborns.

European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology·2014
Same author

Two histone deacetylases, FfHda1 and FfHda2, are important for Fusarium fujikuroi secondary metabolism and virulence.

Applied and environmental microbiology·2013
Same author

Multiple pathways in the oxidation of cis-stilbene by iron-bleomycin.

Journal of the American Chemical Society·2012
Same author

Electroforming of large mirrors.

Applied optics·2010
Same author

In vitro transactivation of Bacillus subtilis RNase P RNA.

FEBS letters·2001
Same author

Photoactivated DNA cleavage by compounds structurally related to the bithiazole moiety of bleomycin.

Bioorganic & medicinal chemistry·2001
Same journal

Correction: Characterization of Mast2 kinase defines structural features, regulation, and substrates.

The Journal of biological chemistry·2026
Same journal

Isotope-Edited ESEEM: A New Method for Probing Copper Binding Sites in Neurodegenerative Proteins.

The Journal of biological chemistry·2026
Same journal

Introduction to the Thematic Review Series on Intracellular Protein Degradation. The ubiquitous biology of intracellular protein degradation: a tribute to Alfred L. ("Fred") Goldberg.

The Journal of biological chemistry·2026
Same journal

Correction: Aromatic residue-rich amino-terminal segments of temporin L self-assemble into collagen-mimetic peptides with cell-adhesion properties.

The Journal of biological chemistry·2026
Same journal

YhbO is a DJ-1 family glyoxalase and α-oxoaldehyde hydratase that confers resistance to reactive carbonyl stress (112).

The Journal of biological chemistry·2026
Same journal

ARMH3 acts as a central scaffold at the Golgi/TGN through interactions with Arl5, GBF1, and PI4KB.

The Journal of biological chemistry·2026
See all related articles

Researchers created dimeric transfer RNA (tRNA) models to study ribonuclease P (RNase P) activity. RNase P and its catalytic subunit M1 RNA cleaved these models, demonstrating the importance of the 3'-half of the tRNA for recognition.

Area of Science:

  • Molecular Biology
  • RNA Biochemistry
  • Enzymology

Background:

  • Ribonuclease P (RNase P) is a crucial enzyme responsible for tRNA maturation.
  • Understanding the substrate recognition mechanism of RNase P is essential for deciphering tRNA processing pathways.
  • Previous studies suggested structural elements in tRNA precursors are important for RNase P cleavage.

Purpose of the Study:

  • To construct and characterize dimeric tRNA molecules as novel substrates for RNase P and M1 RNA.
  • To investigate the role of specific tRNA structural features in the endonucleolytic cleavage reaction.
  • To determine the precise cleavage site within the dimeric tRNA precursor model.

Main Methods:

  • Construction of dimeric tRNA precursor models using T4 RNA ligase-mediated coupling.

Related Experiment Videos

  • Annealing of the 3 omino-half of yeast tRNAPhe to the coupled precursor.
  • Enzymatic cleavage assays using purified Escherichia coli RNase P holoenzyme and M1 RNA.
  • Analysis of cleavage products using polyacrylamide gel electrophoresis and radiolabeling at phosphodiester bonds.

    • Main Results:

    • Both E. coli RNase P and M1 RNA efficiently cleaved the constructed dimeric tRNA precursor models.
    • Cleavage was dependent on the presence of the annealed 3 omino-half of yeast tRNAPhe, indicating its crucial role.
    • The cleavage occurred at the expected phosphate ester linkage, generating products with authentic mobility.
    • radiolabeling confirmed the precise endonucleolytic cleavage site.

    Conclusions:

    • The endonucleolytic separation of two tRNAs by RNase P is significantly influenced by structural features in the 3 omino-half of the 3 omino-tRNA.
    • These findings support the hypothesis that the T stem and loop of the 3 omino-tRNA may act as a recognition site for RNase P.
    • The dimeric tRNA precursor model serves as a valuable tool for studying RNase P substrate recognition and catalytic mechanisms.