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

A complementary DNA oligomer releases a transcription pause complex.

R Fisher, C Yanofsky

    The Journal of Biological Chemistry
    |August 10, 1983
    PubMed
    Summary

    A DNA oligomer prevents RNA hairpin formation, relieving transcription pausing in the tryptophan (trp) operon leader region. This finding suggests a method to disrupt RNA secondary structures that regulate gene expression.

    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

    A novel approach to speed up peptide sequencing via MS/MS spectra analysis.

    Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual Conference·2007
    Same author

    Surface analysis of peptide mass spectra to improve time and mass localization.

    Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual Conference·2007
    Same author

    New insights into regulation of the tryptophan biosynthetic operon in Gram-positive bacteria.

    Trends in genetics : TIG·2005
    Same author

    Inhibition of the B. subtilis regulatory protein TRAP by the TRAP-inhibitory protein, AT.

    Science (New York, N.Y.)·2001
    Same author

    The mechanism of tryptophan induction of tryptophanase operon expression: tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA(Pro).

    Proceedings of the National Academy of Sciences of the United States of America·2001
    Same author

    Advancing our knowledge in biochemistry, genetics, and microbiology through studies on tryptophan metabolism.

    Annual review of biochemistry·2001

    Area of Science:

    • Molecular Biology
    • Gene Regulation
    • Bacterial Transcription

    Background:

    • Expression of tryptophan (trp) operons in bacteria like Escherichia coli is regulated by alternative secondary structures in the trp operon leader transcript.
    • RNA polymerase pauses during in vitro transcription of E. coli trp leader mRNA, specifically near base pair 90, after the formation of a hairpin secondary structure.

    Purpose of the Study:

    • To investigate the role of RNA hairpin formation in transcription pausing.
    • To determine if a DNA oligomer can interfere with RNA hairpin formation and relieve transcription pausing.

    Main Methods:

    • In vitro transcription assays were performed using E. coli trp leader mRNA.
    • A DNA oligomer complementary to the proximal segment of the RNA hairpin was added to observe its effect on transcription pausing.
    • The effect of the DNA oligomer was assessed in the presence and absence of E. coli L-factor, which enhances pausing.

    Main Results:

    • Addition of the DNA oligomer relieved transcription pausing in vitro, both with and without L-factor.
    • The DNA oligomer appears to interfere with the formation of the RNA hairpin, which is recognized by RNA polymerase as a pause signal.
    • The oligomer also relieved pausing in L-factor-induced paused complexes, indicating it can disrupt preformed secondary structures.

    Conclusions:

    • RNA hairpin formation in the trp operon leader transcript is a key signal for transcription pausing.
    • A complementary DNA oligomer can effectively disrupt this RNA hairpin, thereby relieving transcription pausing.
    • This mechanism offers a potential strategy for modulating gene expression by targeting regulatory RNA structures.

    Related Experiment Videos