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

U2 small nuclear RNP assembly in vitro.

A M Kleinschmidt1, J R Patton, T Pederson

  • 1Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, MA 01545.

Nucleic Acids Research
|June 26, 1989
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

Defining ruminal and total-tract starch degradation for adult dairy cattle using in vivo data.

Journal of dairy science·2012
Same author

The molecular cytology of gene expression: fluorescent RNA as both a stain and tracer in vivo.

European journal of histochemistry : EJH·2004
Same author

Human cell lines expressing hormone regulated T7 RNA polymerase localized at distinct intranuclear sites.

Gene·2001
Same author

Is the nucleus in need of translation?

Trends in cell biology·2001
Same author

Protein mobility within the nucleus--what are the right moves?

Cell·2001
Same author

Fluorescent RNA cytochemistry: tracking gene transcripts in living cells.

Nucleic acids research·2001
Same journal

Correction to 'scSuperAnnotator: A platform for benchmarking comparison and visualizing automated cellular annotation methods for scRNA-seq data'.

Nucleic acids research·2026
Same journal

Correction to 'Differentiable partition function calculation for RNA'.

Nucleic acids research·2026
Same journal

Deployment of non-canonical splicing in tunicate genomes is mediated by divergent U2AF function and changing m6A modification in U1 and U6 snRNA.

Nucleic acids research·2026
Same journal

Bacillus subtilis DnaB forms multiple protein-protein interactions essential for DNA replication initiation.

Nucleic acids research·2026
Same journal

Multiple forms of protein-protein and DNA binding are exhibited by BrxC from the BREX phage restriction system.

Nucleic acids research·2026
Same journal

Biosynthesis of glycosylated 5-hydroxycytosine in the DNA of diverse viruses.

Nucleic acids research·2026
See all related articles

ATP is crucial for assembling functional U2 small nuclear ribonucleoprotein complexes (snRNPs). Proper assembly requires ATP hydrolysis and specific U2 RNA sequences, promoting essential RNA modifications like pseudouridine formation.

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biochemistry

Background:

  • U2 small nuclear ribonucleoprotein complexes (snRNPs) are essential for pre-mRNA splicing.
  • The assembly process of U2 snRNPs is not fully understood.
  • Post-transcriptional modifications of U2 RNA play a role in snRNP function.

Purpose of the Study:

  • To investigate the requirements for in vitro assembly of functional U2 snRNPs.
  • To identify the role of ATP and specific RNA sequences in U2 snRNP biogenesis.
  • To examine the impact of assembly conditions on U2 RNA modifications.

Main Methods:

  • In vitro assembly of U2 snRNPs using SP6-transcribed U2 RNA precursor in HeLa cell S100 extracts.
  • Characterization of assembled particles using Cs2SO4 gradients, micrococcal nuclease digestion, and antibody binding assays.

Related Experiment Videos

  • Analysis of U2 RNA sequences and post-transcriptional modifications.

    • Main Results:

    • ATP-dependent assembly yielded particles with properties of native U2 snRNPs, including correct buoyant density and nuclease digestion patterns.
    • Assembled particles reacted with antibodies specific for U2 snRNP proteins (A' and B") and Sm proteins, but not U1 snRNP proteins.
    • Non-hydrolyzable ATP analogs failed to support assembly, indicating ATP hydrolysis is necessary.
    • A specific region (nucleotides 154-167) of U2 RNA was essential for binding U2-specific proteins, but not Sm core proteins.
    • Pseudouridine formation in U2 RNA was enhanced under ATP-dependent assembly conditions.

    Conclusions:

    • ATP hydrolysis is a critical energy-dependent step in the in vitro assembly of functional U2 snRNPs.
    • Specific sequences within U2 RNA are required for the recognition and binding of U2-specific proteins during snRNP assembly.
    • Assembly conditions that support U2 snRNP formation also enhance pseudouridine modification of U2 RNA.