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Verfasst von:Jovanovic, Bogdan [VerfasserIn]   i
 Schubert, Lisa [VerfasserIn]   i
 Poetz, Fabian [VerfasserIn]   i
 Stoecklin, Georg [VerfasserIn]   i
Titel:Tagging of RPS9 as a tool for ribosome purification and identification of ribosome-associated proteins
Verf.angabe:Bogdan Jovanovic, Lisa Schubert, Fabian Poetz, and Georg Stoecklin
Jahr:2021
Umfang:9 S.
Fussnoten:Gesehen am 02.06.2021 ; First published: December 30, 2020
Titel Quelle:Enthalten in: Archives of biological sciences
Ort Quelle:Belgrade : Srpsko Biološko Društvo, 2002
Jahr Quelle:2021
Band/Heft Quelle:73(2021), 1, Seite 47-55
ISSN Quelle:1821-4339
Abstract:Ribosomes, the catalytic machinery required for protein synthesis, are comprised of 4 ribosomal RNAs and about 80 ribosomal proteins in mammals. Ribosomes further interact with numerous associated factors that regulate their biogenesis and function. As mutations of ribosomal proteins and ribosome-associated proteins cause many diseases, it is important to develop tools by which ribosomes can be purified efficiently and with high specificity. Here, we designed a method to purify ribosomes from human cell lines by C-terminally tagging human RPS9, a protein of the small ribosomal subunit. The tag consists of a flag peptide and a streptavidin-binding peptide (SBP) separated by the tobacco etch virus (TEV) protease cleavage site. We demonstrate that RPS9-Flag-TEV-SBP (FTS) is efficiently incorporated into the ribosome without interfering with regular protein synthesis. Using HeLa-GFP-G3BP1 cells stably expressing RPS9-FTS or, as a negative control, mCherry-FTS, we show that complete ribosomes as well as numerous ribosome-associated proteins are efficiently and specifically purified following pull-down of RPS9-FTS using streptavidin beads. This tool will be helpful for the characterization of human ribosome heterogeneity, post-translational modifications of ribosomal proteins, and changes in ribosome-associated factors after exposing human cells to different stimuli and conditions.
DOI:doi:10.2298/ABS20120557J
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext: https://doi.org/10.2298/ABS20120557J
 Volltext: http://www.doiserbia.nb.rs/Article.aspx?ID=0354-46642000057J
 DOI: https://doi.org/10.2298/ABS20120557J
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:affinity purification
 evolution
 panther
 reveals
 ribosome purification
 ribosome-associated proteins
 rps9
 streptavidin-binding peptide
 stress granule
K10plus-PPN:1759442445
Verknüpfungen:→ Zeitschrift

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