SecM-stalled ribosomes adopt an altered geometry at the peptidyl transferase center.
As nascent polypeptide chains are synthesized, they pass through a tunnel in the large ribosomal subunit. Interaction between specific nascent chains and the ribosomal tunnel is used to induce translational stalling for the regulation of gene expression. One well-characterized example is the Escherichia coli SecM (secretion monitor) gene product, which induces stalling to up-regulate translation initiation of the downstream secA gene, which is needed for protein export. Although many of the key components of SecM and the ribosomal tunnel have been identified, understanding of the mechanism by which the peptidyl transferase center of the ribosome is inactivated has been lacking. Here we present a cryo-electron microscopy reconstruction of a SecM-stalled ribosome nascent chain complex at 5.6 Å. While no cascade of rRNA conformational changes is evident, this structure reveals the direct interaction between critical residues of SecM and the ribosomal tunnel. Moreover, a shift in the position of the tRNA-nascent peptide linkage of the SecM-tRNA provides a rationale for peptidyl transferase center silencing, conditional on the simultaneous presence of a Pro-tRNA(Pro) in the ribosomal A-site. These results suggest a distinct allosteric mechanism of regulating translational elongation by the SecM stalling peptide.
Gene Center and Department of Biochemistry, University of Munich, Munich, Germany.
Shashi
S
Bhushan
Thomas
T
Hoffmann
Birgit
B
Seidelt
Jens
J
Frauenfeld
Thorsten
T
Mielke
Otto
O
Berninghausen
Daniel N
DN
Wilson
Roland
R
Beckmann
D020134:Catalytic Domain; D020285:Cryoelectron Microscopy; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D008968:Molecular Conformation; D010458:Peptidyl Transferases / Q000737:chemistry* / Q000378:metabolism; D014176:Protein Biosynthesis; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
D029968:Escherichia coli Proteins; C497347:SecM protein, E coli; D014157:Transcription Factors; D012343:RNA, Transfer; D010458:Peptidyl Transferases
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