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

The yeast U2A'/U2B complex is required for pre-spliceosome formation

F Caspary1, B Séraphin

  • 1EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.

The EMBO Journal
|November 3, 1998
PubMed
Summary

Researchers identified LEA1, a protein similar to human U2A', in yeast. LEA1 and Yib9p are crucial for yeast U2 snRNP function, impacting U2 snRNA levels and pre-mRNA splicing.

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

Strategies for the structural analysis of multi-protein complexes: lessons from the 3D-Repertoire project.

Journal of structural biology·2011
Same author

A new therapeutic approach to treat psoriasis by inhibition of fatty acid oxidation by Etomoxir.

The British journal of dermatology·2005
Same author

The S. cerevisiae SET3 complex includes two histone deacetylases, Hos2 and Hst1, and is a meiotic-specific repressor of the sporulation gene program.

Genes & development·2001
Same author

The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation.

Nucleic acids research·2001
Same author

The tandem affinity purification (TAP) method: a general procedure of protein complex purification.

Methods (San Diego, Calif.)·2001
Same author

Positive feedback in eukaryotic gene networks: cell differentiation by graded to binary response conversion.

The EMBO journal·2001

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Human U2 small nuclear ribonucleoprotein (snRNP) involves proteins U2A' and U2B'' interacting with U2 snRNA.
  • Yeast (Saccharomyces cerevisiae) possesses a U2B'' counterpart, Yib9p, but a U2A' homolog was previously unknown.

Purpose of the Study:

  • To identify and characterize the yeast homolog of human U2A'.
  • To investigate the roles of the identified yeast protein (LEA1) and Yib9p in U2 snRNP assembly and function.
  • To elucidate their impact on pre-mRNA splicing and cell viability.

Main Methods:

  • Database searches to identify potential U2A' homologs.
  • Biochemical assays to confirm Lea1p as a U2 snRNP component and study its interaction with Yib9p.
  • Genetic analysis of yeast strains lacking LEA1 and/or YIB9.
  • In vitro spliceosome assembly assays.
  • Complementation studies using recombinant proteins.

Main Results:

  • A novel yeast protein, LEA1, was identified with high similarity to human U2A'.
  • Lea1p is a specific component of the yeast U2 snRNP and directly interacts with Yib9p.
  • In vivo, Lea1p and Yib9p exhibit interdependent association with U2 snRNA.
  • Deletion of LEA1 and/or YIB9 leads to slow growth, temperature sensitivity, reduced U2 snRNA levels, and impaired pre-mRNA splicing.
  • Spliceosome assembly is blocked before U2 snRNP addition in these mutants.
  • Recombinant Lea1p and Yib9p partially rescue the observed defects.

Conclusions:

  • Lea1p and Yib9p are essential specific proteins for yeast U2 snRNP formation and function.
  • These proteins play a critical role in the early stages of spliceosome assembly, specifically prior to U2 snRNP incorporation.
  • The findings highlight the conserved importance of U2 snRNP-associated proteins in pre-mRNA splicing across species.

Related Experiment Videos