Nascent peptides that block protein synthesis in bacteria

Christopher J. Woolstenhulme, Shankar Parajuli, David W. Healey, Diana P. Valverde, E. Nicholas Petersen, Agata L. Starosta, Nicholas R. Guydosh, W. Evan Johnson, Daniel N. Wilson, Allen R. Buskirk

Research output: Contribution to journalArticle

Abstract

Although the ribosome is a very general catalyst, it cannot synthesize all protein sequences equally well. For example, ribosomes stall on the secretion monitor (SecM) leader peptide to regulate expression of a downstream gene. Using a genetic selection in Escherichia coli, we identified additional nascent peptide motifs that stall ribosomes. Kinetic studies show that some nascent peptides dramatically inhibit rates of peptide release by release factors. We find that residues upstream of the minimal stalling motif can either enhance or suppress this effect. In other stalling motifs, peptidyl transfer to certain aminoacyl-tRNAs is inhibited. In particular, three consecutive Pro codons pose a challenge for elongating ribosomes. The translation factor elongation factor P, which alleviates pausing at polyproline sequences, has little or no effect on other stalling peptides. The motifs that we identified are underrepresented in bacterial proteomes and show evidence of stalling on endogenous E. coli proteins.

Original languageEnglish (US)
Pages (from-to)E878-E887
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number10
DOIs
StatePublished - Mar 5 2013

Keywords

  • EF-P
  • Proline
  • Ribosome stalling
  • TmRNA

ASJC Scopus subject areas

  • General

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  • Cite this

    Woolstenhulme, C. J., Parajuli, S., Healey, D. W., Valverde, D. P., Petersen, E. N., Starosta, A. L., Guydosh, N. R., Johnson, W. E., Wilson, D. N., & Buskirk, A. R. (2013). Nascent peptides that block protein synthesis in bacteria. Proceedings of the National Academy of Sciences of the United States of America, 110(10), E878-E887. https://doi.org/10.1073/pnas.1219536110