We have recently been investigating the regulation of the expression of SecA, a cytoplasmic ATPase that plays a major role in the translocation of proteins through the SecYEG complex in E. coli. The gene that encodes SecA resides immediately downstream of the gene that encodes SecM, and the two genes form a single operon. When secretion is impaired, a 17 amino acid motif (150FXXXXWIXXXXGIRAGP166) near the C-terminus of SecM induces a translation arrest. This motif is recognized inside the ribosome tunnel, but the mechanism of recognition is unknown. While single mutations in the motif impair recognition, we found that novel arrest-inducing peptides can be created through remodeling of the SecM C-terminus. We found that R163 is indispensable, but that flanking residues that vary in number, position, and side chain chemistry play an important secondary role in translation arrest. The observation that individual SecM variants show a distinct pattern of crosslinking to ribosomal proteins suggests that each peptide adopts a unique conformation inside the tunnel. Based on our results, we propose that translation arrest occurs when the peptide conformation specified by flanking residues moves R163 into a precise intra-tunnel location. Our data indicate that translation arrest results from extensive communication between SecM and the ribosome tunnel and help explain the striking diversity of arrest-inducing peptides found in bacteria, fungi and higher eukaryotes.
| Yap, Mee-Ngan F; Bernstein, Harris D (2013) Mutations in the Escherichia coli ribosomal protein L22 selectively suppress the expression of a secreted bacterial virulence factor. J Bacteriol 195:2991-9 |
| Yap, Mee-Ngan; Bernstein, Harris D (2011) The translational regulatory function of SecM requires the precise timing of membrane targeting. Mol Microbiol 81:540-53 |
| Peterson, Janine H; Woolhead, Cheryl A; Bernstein, Harris D (2010) The conformation of a nascent polypeptide inside the ribosome tunnel affects protein targeting and protein folding. Mol Microbiol : |
| Yap, Mee-Ngan; Bernstein, Harris D (2009) The plasticity of a translation arrest motif yields insights into nascent polypeptide recognition inside the ribosome tunnel. Mol Cell 34:201-11 |
