Nachbarschaftsstudien am Ribosom: proximity dependent biotin identification

Smolinski, Nadine; Valerius, Oliver

doi:10.1007/s12268-016-0665-4

Cited by 1 publication

(1 citation statement)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The resulting proteome values were used to normalize affinity capture SILAC ratios against total protein abundances. Modified from Smolinski and Valerius (74). B, All four BioID-experiments were performed with triple SILAC to quantitatively compare cells of three different cultures in one batch.…”

Section: Resultsmentioning

confidence: 99%

Capturing the Asc1p/Receptor for Activated C Kinase 1 (RACK1) Microenvironment at the Head Region of the 40S Ribosome with Quantitative BioID in Yeast

Opitz

Schmitt

Hofer-Pretz

et al. 2017

Molecular & Cellular Proteomics

Self Cite

101

View full text Add to dashboard Cite

The Asc1 protein of is a scaffold protein at the head region of ribosomal 40S that links mRNA translation to cellular signaling. In this study, proteins that colocalize with Asc1p were identified with proximity-dependenttin entification (BioID), an labeling technique described here for the first time for yeast. Biotinylated Asc1p-birA*-proximal proteins were identified and quantitatively verified against controls applying SILAC and mass spectrometry. The mRNA-binding proteins Sro9p and Gis2p appeared together with Scp160p, each providing ribosomes with nuclear transcripts. The cap-binding protein eIF4E (Cdc33p) and the eIF3/a-subunit (Rpg1p) were identified reflecting the encounter of proteins involved in the initiation of mRNA translation at the head region of ribosomal 40S. Unexpectedly, a protein involved in ribosome preservation (the clamping factor Stm1p), the deubiquitylation complex Ubp3p-Bre5p, the RNA polymerase II degradation factor 1 (Def1p), and transcription factors (Spt5p, Mbf1p) colocalize with Asc1p in exponentially growing cells. For Asc1p, a variant considered to be deficient in binding to ribosomes, BioID revealed its predominant ribosome localization. Glucose depletion replaced most of the Asc1p colocalizing proteins for additional ribosomal proteins, suggesting a ribosome aggregation process during early nutrient limitation, possibly concomitant with ribosomal subunit clamping. Overall, the characterization of the Asc1p microenvironment with BioID confirmed and substantiated our recent findings that the β-propeller broadly contributes to signal transduction influencing phosphorylation of colocalizing proteins ( of Bre5p), and by that might affect nuclear gene transcription and the fate of ribosomes.

show abstract

Section: Resultsmentioning

confidence: 99%