The signal recognition particle binds to protein L23 at the peptide exit of the Escherichia coli ribosome

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Date
2003
Authors
Gu, Shan-Qing
Peske, Frank
Wieden, Hans-Joachim
Rodnina, Marina V.
Wintermeyer, Wolfgang
Journal Title
Journal ISSN
Volume Title
Publisher
Cold Springs Harbor Laboratory Press
Abstract
The signal recognition particle (SRP) from Escherichia coli, composed of Ffh protein and 4.5S RNA, mediates membrane targeting of translating ribosomes displaying a signal or signal-anchor sequence. SRP binds at the peptide exit of the large ribosomal subunit. Structural details of the interaction are not known. Here, the position of Ffh or SRP on the ribosome was probed by using site-specific UV-induced crosslinking by p-azidophenacyl bromide (AzP) attached to a number of cysteine residues engineered into surface positions of Ffh. Efficient crosslinking to vacant ribosomes took place from two positions (AzP17 and AzP25) in the N domain of Ffh, both with Ffh and SRP. Both AzP17 and AzP25 were predominantly crosslinked to ribosomal protein L23 that is located at the peptide exit of the 50S subunit.The SRP receptor, FtsY, did not change the crosslink pattern, whereas the presence of a nascent signal peptide on the ribosome resulted in a second crosslink between Ffh(AzP17) and protein L23, indicating that binding to the nascent signal peptide induced a slightly different arrangement of SRP on the ribosome. These results indicate a model of the topographical arrangement of SRP at the peptide exit of the 50S ribosomal subunit.
Description
Sherpa Romeo green journal. Permission to archive final published version
Keywords
4.5S RNA , Membrane targeting , Ribosome nascent chain complexes , Signal peptide , Crosslinking (Polymerization) , Protein L23
Citation
Gu, S., Peske, F., Wieden, H. J., Rodnina, M. V., & Wintermeyer, W. (2003). The signal recognition particle binds to protein L23 at the peptide exit of the Escherichia coli ribosome. RNA, 9(5), 566-73. http://www.rnajournal.org/cgi/doi/ 10.1261/rna.2196403.