TCBG Seminar

Nascent peptide sequences that stall ribosomes during protein synthesis

Professor Allen Buskirk
Department of Chemistry and Biochemistry
Brigham Young University
Provo, Utah

Monday, February 14, 2011
3:00 pm (CT)
3269 Beckman Institute


Several nascent peptides stall ribosomes during their own translation in both prokaryotes and eukaryotes. Leader peptides that induce stalling can regulate downstream gene expression. Interestingly, stalling peptides show little sequence similarity and interact with the ribosome through distinct mechanisms. To explore the scope of regulation by stalling peptides and to better understand the mechanism of stalling, we identified and characterized new examples from random libraries. We created a genetic selection that ties the life of Escherichia coli cells to stalling at a specific site. This selection relies on the natural bacterial system that rescues arrested ribosomes. We altered transfer-messenger RNA, a key component of this rescue system, to direct the completion of a necessary protein if and only if stalling occurs. We identified three classes of stalling peptides: C- terminal Pro residues, SecM-like peptides, and the novel stalling sequence FxxYxIWPP. Like the leader peptides SecM and TnaC, the FxxYxIWPP peptide induces stalling efficiently by inhibiting peptidyl transfer. The nascent peptide exit tunnel and peptidyl-transferase center are implicated in this stalling event, although mutations in the ribosome affect stalling on SecM and FxxYxIWPP differently. We conclude that ribosome stalling can be caused by numerous sequences and is more common than previously believed.

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