During translation, the first encounter of nascent polypeptides is with the ribosome-associated chaperones that assist the folding process—a principle that seems to be conserved in evolution1,2,3. In Escherichia coli, the ribosome-bound Trigger Factor chaperones the folding of cytosolic proteins by interacting with nascent polypeptides4,5. Here we identify a ribosome-binding motif in the amino-terminal domain of Trigger Factor. We also show the formation of crosslinked products between Trigger Factor and two adjacent ribosomal proteins, L23 and L29, which are located at the exit of the peptide tunnel in the ribosome. L23 is essential for the growth of E. coli and the association of Trigger Factor with the ribosome, whereas L29 is dispensable in both processes. Mutation of an exposed glutamate in L23 prevents Trigger Factor from interacting with ribosomes and nascent chains, and causes protein aggregation and conditional lethality in cells that lack the protein repair function of the DnaK chaperone. Purified L23 also interacts specifically with Trigger Factor in vitro. We conclude that essential L23 provides a chaperone docking site on ribosomes that directly links protein biosynthesis with chaperone-assisted protein folding.
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We thank the members of the Bukau lab for discussions; T. Hesterkamp for suggesting the TF FRK/AAA mutation; K. Turgay for suggesting the S-tag experiment; and D. Dougan for comments on the manuscript. This work was supported by grants of the Deutsche Forschungsgemeinschaft to B.B. and E.D, the Human Frontier Science Program to E.D. and N.B., the Swiss National Science Foundation to N.B., and fellowships of the Boehringer Ingelheim Fonds to T.R. and the Fonds der Chemischen Industrie (Kékulé scholarship) to W.R.
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Kramer, G., Rauch, T., Rist, W. et al. L23 protein functions as a chaperone docking site on the ribosome. Nature 419, 171–174 (2002) doi:10.1038/nature01047
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