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Naf1 p is a box H/ACA snoRNP assembly factor
Alessandro Fatica1, Mensur Dlakić, David Tollervey
1Wellcome Trust Centre for Cell Biology, Swann Building, King's Buildings, University of Edinburgh, Edinburgh EH9 3JR, UK.
RNA (New York, N.Y.)
|January 8, 2003
Summary
A novel protein, Naf1p, is essential for the accumulation of box H/ACA small nucleolar RNAs (snoRNAs). Naf1p binds to nascent snoRNAs and promotes their assembly into functional small nucleolar ribonucleoprotein particles (snoRNPs).
Area of Science:
- Molecular Biology
- RNA Biology
- Gene Regulation
Background:
- Box H/ACA small nucleolar ribonucleoprotein particles (snoRNPs) are crucial for processing and modifying specific RNA molecules.
- Four proteins (Cbf5p, Gar1p, Nhp2p, Nop10p) are essential for H/ACA snoRNP function, with most required for snoRNA accumulation.
Purpose of the Study:
- To identify novel proteins involved in box H/ACA snoRNP biogenesis.
- To characterize the function and interactions of a newly discovered essential protein, Naf1p.
Main Methods:
- Bioinformatic database searches to identify potential novel proteins.
- Yeast two-hybrid assays to investigate protein-protein interactions.
- In vitro RNA-binding assays.
- Analysis of protein localization and its effect on snoRNA accumulation.
Main Results:
- A novel essential protein, Naf1p, was identified with homology to RNA-binding proteins.
- Naf1p is localized to the nucleus and is required for the accumulation of all box H/ACA snoRNAs, independent of transcription initiation or termination.
- Naf1p exhibits in vitro RNA-binding activity and interacts with Cbf5p and Nhp2p.
- Naf1p binds to the phosphorylated C-terminal domain (CTD) of RNA polymerase II in vivo.
Conclusions:
- Naf1p is a novel essential factor in box H/ACA snoRNP biogenesis.
- Naf1p is recruited to RNA polymerase II via its CTD and binds nascent box H/ACA snoRNAs.
- This interaction facilitates snoRNP assembly and promotes snoRNA accumulation.