Role of Has1 RNA helicase in ribosome biogenesis in yeast S. cerevisiae
The RNA helicase Has1 is involved in the biogenesis of both small and large ribosomal subunit. How it performs these separate roles is not fully understood. We analysed a detailed composition of RNA-protein complexes purified via Has1.
Sample Processing Protocol
Strains expressing Has1 fused to the FTP tandem affinity tag (FLAG-TEV protease site-protein A) were grown in heavy or light media (SILAC). An exponentialy growing cultures were harvested, lysed and Has1 was affinity purified using both tandem affinity tags. The purified complexes were run shortly into a SDS PAGE gel and slices of the gel were submitted for mass spec analysis(in gel digestion) using Orbitrap.
Data Processing Protocol
The obtained Raw data was processed using MaxQuant software. The parameters can be found in the provided files.
Martin Kos, University of Heidelberg, Biochemistry Centre (BZH)
Im Neuenheimer Feld 328
Martin Koš, University of Heidelberg Biochemistry Center Im Neuenheimer Feld 328 69120 Heidelberg Germany ( lab head )
Gnanasundram SV, Kos-Braun IC, Koš M. At least two molecules of the RNA helicase Has1 are simultaneously present in pre-ribosomes during ribosome biogenesis. Nucleic Acids Res. 2019 PubMed: 31511893