tRNA/mRNA Mimicry by tmRNA and SmpB in<i>Trans</i>-Translation

Kurita, Daisuke; Muto, Akira; Himeno, Hyouta

doi:10.4061/2011/130581

Cited by 5 publications

(3 citation statements)

References 79 publications

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“…41,49 This interaction occurs at pre-accommodation and it triggers the accommodation of tmRNA into the A-site after GTP hydrolysis by EF-Tu, compensating for the lack of a codon during the entering of tmRNA while the upper half of tRNA is mimicked by the tRNA domain (TLD). 50 The positioning of SmpB into the decoding site is consistent with the crystal structures of the protein bound to the TLD. 12,13 Overall, the 'RNA-protein' complex mimics the L-shaped conformation of a canonical tRNA.…”

Section: O N O T D I S T R I B U T Esupporting

confidence: 59%

SmpB as the handyman of tmRNA during trans-translation

Felden¹,

Gillet²

2011

RNA Biology

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Section: O N O T D I S T R I B U T Esupporting

confidence: 59%

SmpB as the handyman of tmRNA during trans-translation

Felden¹,

Gillet²

2011

RNA Biology

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“…In our ongoing research on trans-translation, we were trying to produce the A. aeolicus tmRNA using the pBSTNAV vector in E. coli. Trans-translation is a highly sophisticated process in bacteria that recycles ribosomes stalled on defective mRNAs and adds a short tag-peptide to the C-terminus of the incomplete polypeptide as degradation signal [for review, see (48)]. The process of trans-translation uses the tmRNA, which is a unique molecule with dual tRNA and mRNA functions ( Figure 4A).…”

Section: Ms2-encapsulated Rna Is Protected From Ribonucleases: Examplmentioning

confidence: 99%

Co-expression of RNA–protein complexes in Escherichia coli and applications to RNA biology

Ponchon

Catala

Seijo

et al. 2013

Nucleic Acids Research

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RNA has emerged as a major player in many cellular processes. Understanding these processes at the molecular level requires homogeneous RNA samples for structural, biochemical and pharmacological studies. We previously devised a generic approach that allows efficient in vivo expression of recombinant RNA in Escherichia coli. In this work, we have extended this method to RNA/protein co-expression. We have engineered several plasmids that allow overexpression of RNA–protein complexes in E. coli. We have investigated the potential of these tools in many applications, including the production of nuclease-sensitive RNAs encapsulated in viral protein pseudo-particles, the co-production of non-coding RNAs with chaperone proteins, the incorporation of a post-transcriptional RNA modification by co-production with the appropriate modifying enzyme and finally the production and purification of an RNA–His-tagged protein complex by nickel affinity chromatography. We show that this last application easily provides pure material for crystallographic studies. The new tools we report will pave the way to large-scale structural and molecular investigations of RNA function and interactions with proteins.

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“…In vivo and in vitro studies have revealed the significance of the sequence between PK1 and the tag-encoding region, especially at positions –6 to +1 ( Williams et al, 1999 ; Lee et al, 2001 ). In vitro trans -translation system can identify not only the efficiency of the first peptidyl-transfer of trans -translation but also the second amino acid of the tag-peptide, which enables us to specify the start point of the tag-translation ( Lee et al, 2001 ; Konno et al, 2007 ; Takada et al, 2007 ; Kurita et al, 2011 ). Using this system, we have found that some point mutations within the span of -6 to +1 reduce the trans -translation efficiency and/or shift the resuming point of translation on tmRNA by -1 or +1, indicating that this region serves not only as the enhancer of trans -translation but also as the determinant for the resume codon for the tag-peptide ( Lee et al, 2001 ; Konno et al, 2007 ).…”

Section: Determination Of the Resume Codonmentioning

confidence: 99%

Mechanism of trans-translation revealed by in vitro studies

2014

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tmRNA is a bacterial small RNA having a structure resembling the upper half of tRNA and its 3′ end accepts alanine followed by binding to EF-Tu like tRNA. Instead of lacking a lower half of the cloverleaf structure including the anticodon, tmRNA has a short coding sequence for tag-peptide that serves as a target of cellular proteases. An elaborate coordination of two functions as tRNA and mRNA facilitates an irregular translation termed trans-translation: a single polypeptide is synthesized from two mRNA molecules. It allows resumption of translation stalled on a truncated mRNA, producing a chimeric polypeptide comprising the C-terminally truncated polypeptide derived from truncated mRNA and the C-terminal tag-peptide encoded by tmRNA. Trans-translation promotes recycling of the stalled ribosomes in the cell, and the resulting C-terminally tagged polypeptide is preferentially degraded by cellular proteases. Biochemical studies using in vitro trans-translation systems together with structural studies have unveiled the molecular mechanism of trans-translation, during which the upper and lower halves of tRNA are mimicked by the tRNA-like structure of tmRNA and a tmRNA-specific binding protein called SmpB, respectively. They mimic not only the tRNA structure but also its behavior perhaps at every step of the trans-translation process in the ribosome. Furthermore, the C-terminal tail of SmpB, which is unstructured in solution, occupies the mRNA path in the ribosome to play a crucial role in trans-translation, addressing how tmRNA·SmpB recognizes the ribosome stalled on a truncated mRNA.

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tRNA/mRNA Mimicry by tmRNA and SmpB inTrans-Translation

Cited by 5 publications

References 79 publications

SmpB as the handyman of tmRNA during trans-translation

SmpB as the handyman of tmRNA during trans-translation

Co-expression of RNA–protein complexes in Escherichia coli and applications to RNA biology

Mechanism of trans-translation revealed by in vitro studies

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