A structure of a ribosome stalled during translation of the SecM peptide provides insight into the mechanism by which the large subunit active site is inactivated

B Schuwirth

B Seidelt

Birgit Seidelt

C Bernabeu

C Deutsch

C. A Woolhead

D Moazed

D Vasquez

D. R Morris

D. R Tanner

Daniel N. Wilson

E. F Pettersen

F Gong

Gunter Blobel

H Muto

H Nakatogawa

J Frank

J Harms

J Lu

J. A Mindell

J. L Hansen

Jens Frauenfeld

K Bergemann

K Fredrick

K Ito

K Mitra

L Cruz-Vera

L. B Jenner

M Halic

M Selmer

M Simonovic

M. E Butkus

M. G Lawrence

M. N Yap

M. Y Pavlov

N Ban

N Polacek

N Vazquez-Laslop

Otto Berninghausen

P Emsley

P Julian

P Nissen

P. A Penczek

P. S Lovett

R. K Tompkins

Roland Beckmann

S Bhushan

S Fulle

S. C Blanchard

Shashi Bhushan

T Becker

T Tenson

T Wagenknecht

T. M Schmeing

Thomas Hoffmann

Thorsten Mielke

W Humphrey

X Agirrezabala

English

PubMed

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 A. 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

Bhushan, S.

Hoffmann, T.

Seidelt, B.

Frauenfeld, J.

Mielke, T.

Berninghausen, O.

Wilson, D.

Beckmann, R.

MPG.PuRe

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

Bhushan, Shashi

Hoffmann, Thomas

Seidelt, Birgit

Frauenfeld, Jens

Mielke, Thorsten

Berninghausen, Otto

Wilson, Daniel N

Beckmann, Roland

Open Access LMU ( Ludwig-Maximilians-Univ. München)

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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-tRNAPro in the ribosomal A-site. These results suggest a distinct allosteric mechanism of regulating translational elongation by the SecM stalling peptide

CiteSeerX

Crossref

PLoS Biology

SecM-Stalled Ribosomes Adopt an Altered Geometry at the Peptidyl Transferase Center

Universität München: Elektronischen Publikationen

Bhushan Shashi

Hoffmann Thomas

Seidelt Birgit

Frauenfeld Jens

Mielke Thorsten

Berninghausen Otto

Wilson Daniel N.

Beckmann Roland

Public Library of Science (PLOS)

Daniel N Wilson

Directory of Open Access Journals

SecM-stalled ribosomes adopt an altered geometry at the peptidyl transferase center.

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SecM-Stalled Ribosomes Adopt an Altered Geometry at the Peptidyl Transferase Center

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MPG.PuRe

Open Access LMU ( Ludwig-Maximilians-Univ. München)

CiteSeerX

Crossref

Universität München: Elektronischen Publikationen

Public Library of Science (PLOS)

Directory of Open Access Journals

MPG.PuRe