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Related Experiment Videos

Interaction between plasminogen activator inhibitor type-2 and pre-mRNA processing factor 8.

Jing Fan1, Yu-Qing Zhang, Ping Li

  • 1Department of Molecular Genetics, Shanghai Medical School of Fudan University, Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai 200032, China.

Acta Biochimica Et Biophysica Sinica
|September 4, 2004
PubMed
Summary

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Plasminogen activator inhibitor type-2 (PAI-2) protects against apoptosis via a specific fragment that interacts with pre-mRNA processing factor 8 (PRPF8). This interaction was confirmed in vivo but appears unrelated to the apoptosis process itself.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Protein Interactions

Background:

  • Plasminogen activator inhibitor type-2 (PAI-2) is known to protect cells from apoptosis.
  • A specific 33-amino acid fragment between helices C and D of PAI-2 is implicated in this protective function.
  • This fragment is hypothesized to interact with unknown intracellular proteins.

Purpose of the Study:

  • To identify intracellular proteins interacting with the apoptosis-protective fragment of PAI-2.
  • To investigate the in vivo interaction between PAI-2 and its binding partners.
  • To determine if the identified interaction is involved in the PAI-2-mediated apoptosis protection.

Main Methods:

  • Yeast two-hybrid screening using the PAI-2 C-D helix fragment as bait.

Related Experiment Videos

  • Construction of a HeLa cell cDNA library from apoptotic cells.
  • Co-immunoprecipitation assays to confirm in vivo protein interactions.

    • Main Results:

    • A yeast two-hybrid screen identified a 94-amino acid C-terminal fragment of pre-mRNA processing factor 8 (PRPF8) as an interacting partner of the PAI-2 fragment.
    • Co-immunoprecipitation confirmed that PAI-2 interacts with PRPF8 in vivo.
    • PAI-2 was shown to bind the C-terminus of PRPF8 both intracellularly and extracellularly.

    Conclusions:

    • The C-terminal fragment of PAI-2 interacts with PRPF8.
    • This interaction occurs in vivo and is not restricted to the nucleus.
    • The findings suggest that the PAI-2 and PRPF8 interaction may not play a direct role in PAI-2's apoptosis-protective mechanism.