Structural basis for translational stalling by human cytomegalovirus and fungal arginine attenuator peptide

Shashi Bhushan; Helge Meyer; Agata L Starosta; Thomas Becker; Thorsten Mielke; Otto Berninghausen; Michael Sattler; Daniel N Wilson; Roland Beckmann

doi:10.1016/j.molcel.2010.09.009

Structural basis for translational stalling by human cytomegalovirus and fungal arginine attenuator peptide

Mol Cell. 2010 Oct 8;40(1):138-46. doi: 10.1016/j.molcel.2010.09.009.

Authors

Shashi Bhushan¹, Helge Meyer, Agata L Starosta, Thomas Becker, Thorsten Mielke, Otto Berninghausen, Michael Sattler, Daniel N Wilson, Roland Beckmann

Affiliation

¹ Gene Center and Department of Biochemistry and Center for integrated Protein Science Munich, University of Munich, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.

PMID: 20932481
DOI: 10.1016/j.molcel.2010.09.009

Abstract

Specific regulatory nascent chains establish direct interactions with the ribosomal tunnel, leading to translational stalling. Despite a wealth of biochemical data, structural insight into the mechanism of translational stalling in eukaryotes is still lacking. Here we use cryo-electron microscopy to visualize eukaryotic ribosomes stalled during the translation of two diverse regulatory peptides: the fungal arginine attenuator peptide (AAP) and the human cytomegalovirus (hCMV) gp48 upstream open reading frame 2 (uORF2). The C terminus of the AAP appears to be compacted adjacent to the peptidyl transferase center (PTC). Both nascent chains interact with ribosomal proteins L4 and L17 at tunnel constriction in a distinct fashion. Significant changes at the PTC were observed: the eukaryotic-specific loop of ribosomal protein L10e establishes direct contact with the CCA end of the peptidyl-tRNA (P-tRNA), which may be critical for silencing of the PTC during translational stalling. Our findings provide direct structural insight into two distinct eukaryotic stalling processes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor / biosynthesis
Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor / chemistry*
Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor / genetics
Circular Dichroism
Cryoelectron Microscopy
Cytomegalovirus / genetics
Cytomegalovirus / metabolism*
Gene Expression Regulation, Fungal
Magnetic Resonance Spectroscopy
Models, Molecular
Nucleic Acid Conformation
Open Reading Frames
Peptide Fragments / biosynthesis
Peptide Fragments / chemistry*
Peptide Fragments / genetics
Peptidyl Transferases / chemistry
Protein Biosynthesis*
Protein Conformation
RNA, Transfer, Amino Acyl / chemistry
Ribosomal Proteins / chemistry
Ribosomes / metabolism
Ribosomes / ultrastructure*
Structure-Activity Relationship
Viral Envelope Proteins / biosynthesis
Viral Envelope Proteins / chemistry*
Viral Envelope Proteins / genetics
Yeasts / genetics
Yeasts / metabolism*

Substances

Peptide Fragments
RNA, Transfer, Amino Acyl
Ribosomal Proteins
UL4 protein, Human cytomegalovirus
Viral Envelope Proteins
arginine attenuator peptide
ribosomal protein L4
tRNA, peptidyl-
Peptidyl Transferases
Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor