Pech M.,Abteilung Vingron AG |
Nierhaus K.H.,Abteilung Vingron AG |
Nierhaus K.H.,Charite - Medical University of Berlin
Molecular Microbiology | Year: 2012
The tmRNA/SmpB system, which is almost universal in bacteria, rescues bacterial ribosomes stalled at the end of non-stop mRNAs (mRNAs lacking a stop codon). In addition, a few bacteria, including Escherichia coli, have developed a second two-component system as reported by Chadani etal. A small protein, ArfA of 55 amino acids (formerly called YdhL), mediates binding of release factor 2 to the ribosomal A site lacking a complete mRNA codon and thereby triggers translational termination and rescue of the stalled ribosome. © 2012 Blackwell Publishing Ltd.
Zhang D.,CAS Institute of Biophysics |
Liu G.,CAS Institute of Biophysics |
Xue J.,CAS Institute of Biophysics |
Lou J.,CAS Institute of Biophysics |
And 4 more authors.
Nucleic Acids Research | Year: 2012
Translational GTPases (trGTPases) regulate all phases of protein synthesis. An early event in the interaction of a trGTPase with the ribosome is the contact of the G-domain with the C-terminal domain (CTD) of ribosomal protein L12 (L12-CTD) and subsequently interacts with the N-terminal domain of L11 (L11-NTD). However, the structural and functional relationships between L12-CTD and L11-NTD remain unclear. Here, we performed mutagenesis, biochemical and structural studies to identify the interactions between L11-NTD and L12-CTD. Mutagenesis of conserved residues in the interaction site revealed their role in the docking of trGTPases. During docking, loop62 of L11-NTD protrudes into a cleft in L12-CTD, leading to an open conformation of this domain and exposure of hydrophobic core. This unfavorable situation for L12-CTD stability is resolved by a chaperone-like activity of the contacting G-domain. Our results suggest that all trGTPases-regardless of their different specific functions-use a common mechanism for stabilizing the L11-NTD.L12-CTD interactions. © 2012 The Author(s).
Budkevich T.,Charite - Medical University of Berlin |
Budkevich T.,Abteilung Vingron AG |
Budkevich T.,NASU Institute of Molecular Biology and Genetics |
Giesebrecht J.,Charite - Medical University of Berlin |
And 7 more authors.
Molecular Cell | Year: 2011
Although the structural core of the ribosome is conserved in all kingdoms of life, eukaryotic ribosomes are significantly larger and more complex than their bacterial counterparts. The extent to which these differences influence the molecular mechanism of translation remains elusive. Multiparticle cryo-electron microscopy and single-molecule FRET investigations of the mammalian pretranslocation complex reveal spontaneous, large-scale conformational changes, including an intersubunit rotation of the ribosomal subunits. Through structurally related processes, tRNA substrates oscillate between classical and at least two distinct hybrid configurations facilitated by localized changes in their L-shaped fold. Hybrid states are favored within the mammalian complex. However, classical tRNA positions can be restored by tRNA binding to the E site or by the eukaryotic-specific antibiotic and translocation inhibitor cycloheximide. These findings reveal critical distinctions in the structural and energetic features of bacterial and mammalian ribosomes, providing a mechanistic basis for divergent translation regulation strategies and species-specific antibiotic action. © 2011 Elsevier Inc.
Budkevich T.V.,Charite - Medical University of Berlin |
Budkevich T.V.,Abteilung Vingron AG |
Budkevich T.V.,NASU Institute of Molecular Biology and Genetics |
Giesebrecht J.,Charite - Medical University of Berlin |
And 12 more authors.
Cell | Year: 2014
The extent to which bacterial ribosomes and the significantly larger eukaryotic ribosomes share the same mechanisms of ribosomal elongation is unknown. Here, we present subnanometer resolution cryoelectron microscopy maps of the mammalian 80S ribosome in the posttranslocational state and in complex with the eukaryotic eEF1A·Val-tRNA·GMPPNP ternary complex, revealing significant differences in the elongation mechanism between bacteria and mammals. Surprisingly, and in contrast to bacterial ribosomes, a rotation of the small subunit around its long axis and orthogonal to the well-known intersubunit rotation distinguishes the posttranslocational state from the classical pretranslocational state ribosome. We term this motion "subunit rolling." Correspondingly, a mammalian decoding complex visualized in substates before and after codon recognition reveals structural distinctions from the bacterial system. These findings suggest how codon recognition leads to GTPase activation in the mammalian system and demonstrate that in mammalia subunit rolling occurs during tRNA selection. © 2014 Elsevier Inc.
Muhs M.,Charite - Medical University of Berlin |
Yamamoto H.,Niigata University |
Yamamoto H.,Abteilung Vingron AG |
Ismer J.,Charite - Medical University of Berlin |
And 10 more authors.
Nucleic Acids Research | Year: 2011
Some viruses exploit internal initiation for their propagation in the host cell. This type of initiation is facilitated by structured elements (internal ribosome entry site, IRES) upstream of the initiator AUG and requires only a reduced number of canonical initiation factors. An important example are IRES of the virus family Dicistroviridae that bind to the inter-subunit side of the small ribosomal 40S subunit and lead to the formation of elongation-competent 80S ribosomes without the help of any initiation factor. Here, we present a comprehensive functional and structural analysis of eukaryotic-specific ribosomal protein rpS25 in the context of this type of initiation and propose a structural model explaining the essential involvement of rpS25 for hijacking the ribosome. © 2011 The Author(s).