Structure of the human 20S U5 snRNP
Schneider, S., Brandina, I., Peter, D. et al. Structure of the human 20S U5 snRNP. Nat Struct Mol Biol 31, 752–756 (2024).
Credits for Summary: Khyati Shukla Aakash Khurana
The study focuses on the molecular interactions of CD2BP2, a protein involved in pre-mRNA splicing, particularly its role in the assembly of the U4/U6.U5 tri-snRNP complex. The researchers employed a combination of biochemical assays, cryo-electron microscopy (cryo-EM), and mass spectrometry to elucidate the structural details and functional implications of CD2BP2 in the spliceosome assembly process.
HEK293T cells were utilized for the PRP6-CD2BP2 pull-down experiment, where plasmids encoding specific protein domains were transfected for interaction studies. Cryo-EM data was collected and processed, followed by structural analysis and atomic modeling using cryo-EM density maps and AlphaFold2 predictions. Additionally, TMT-plex quantitative mass spectrometry was employed for analyzing the eluates from the pull-down experiments.
The study revealed that the CD2BP2GYF domain competes with CD2BP2D1 for binding to PRP8, shedding light on the regulatory mechanisms of spliceosome assembly. Furthermore, CD2BP2 was found to facilitate the recruitment of PRP6 and DIM1, crucial components of the tri-snRNP complex, highlighting its role in spliceosome maturation. These findings provide valuable insights into the molecular mechanisms underlying pre-mRNA splicing and offer a foundation for further investigations into the biogenesis and recycling of the U5 snRNP complex.
The structural and functional characterization of CD2BP2 in the spliceosome assembly process not only enhances our understanding of pre-mRNA splicing but also opens up avenues for exploring potential therapeutic targets related to splicing dysregulation in diseases. The study's comprehensive approach combining biochemical, structural, and proteomic techniques underscores the importance of multidisciplinary strategies in unraveling complex biological processes.