Human prefoldin modulates co-transcriptional pre-mRNA splicing.

Area of Science:

• Molecular Biology
• Genetics
• Cell Biology

Background:

• Prefoldin is a conserved protein complex involved in protein folding.
• Prefoldin has known roles in transcription and chromatin dynamics in yeast.
• Its specific functions in human gene regulation remain incompletely understood.

Purpose of the Study:

• To investigate the role of human prefoldin in genome-wide gene expression and RNA splicing.
• To elucidate the molecular mechanisms by which prefoldin influences transcription and splicing.

Main Methods:

• Genome-wide analysis of gene expression and prefoldin binding.
• Assessment of pre-mRNA splicing efficiency and co-transcriptional splicing.
• Analysis of RNA polymerase II C-terminal domain (CTD) phosphorylation and associated kinase recruitment.

Main Results:

• Prefoldin depletion causes widespread transcriptional alterations and severe pre-mRNA splicing defects in human cells.
• Co-transcriptional splicing is impaired, particularly affecting long genes with many introns.
• Prefoldin loss reduces RNA polymerase II CTD phosphorylation (Ser2 and Ser5) and recruitment of CDK9, impacting splicing factor association.

Conclusions:

• Human prefoldin directly modulates gene expression by affecting the phosphorylation of elongating RNA polymerase II.
• Prefoldin regulates co-transcriptional splicing through its influence on RNA polymerase II CTD.
• These findings reveal a novel role for prefoldin in coordinating transcription and splicing in humans.