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Related Experiment Videos

U snRNP assembly in yeast involves the La protein.

D Xue1, D A Rubinson, B K Pannone

  • 1Departments of Cell Biology and Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06536, USA.

The EMBO Journal
|April 4, 2000
PubMed
Summary

The La protein (Lhp1p) is crucial for stabilizing RNA polymerase III transcripts and also aids in forming RNA polymerase II-transcribed ribonucleoprotein complexes (RNPs), particularly U4/U6 snRNPs, for yeast cell viability.

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Eukaryotic Gene Expression

Background:

  • The La autoantigen is known to bind nascent RNA polymerase III transcripts, protecting them from degradation.
  • Ribonucleoprotein complexes (RNPs) are essential for various cellular processes, including RNA processing and gene regulation.

Purpose of the Study:

  • To investigate the role of the La protein (Lhp1p) in the assembly of RNA polymerase II-transcribed RNAs into RNPs.
  • To understand the function of Lhp1p in spliceosomal small nuclear ribonucleoprotein (snRNP) biogenesis.

Main Methods:

  • Yeast genetics: Utilizing a temperature-sensitive mutant (smd1-1) affecting spliceosomal protein Smd1p.
  • RNA immunoprecipitation and Northern blotting to detect and quantify specific snRNAs and snRNPs.
  • Analysis of Lhp1p binding to pre-snRNAs in vitro.

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Main Results:

  • Lhp1p is required for yeast cell growth at low temperatures, especially in the smd1-1 mutant background.
  • Lhp1p binds to precursors of U1, U2, U4, and U5 RNAs in both wild-type and mutant yeast cells.
  • Lhp1p is essential for the accumulation of U4/U6 snRNPs and cell viability at low temperatures, facilitating Sm protein binding to pre-U4 RNA.

Conclusions:

  • The La protein (Lhp1p) plays a dual role, stabilizing RNA polymerase III transcripts and assisting in the formation of specific RNA polymerase II-transcribed RNPs.
  • Lhp1p is critical for the efficient biogenesis of U4/U6 snRNPs by stabilizing a 3'-extended form of U4 RNA, thereby promoting Sm protein incorporation.
  • This function of Lhp1p is vital for spliceosome assembly and cell viability under specific stress conditions.