Related Experiment Video

Updated: Aug 31, 2026

Xenopus laevis as a Model to Identify Translation Impairment
10:24

Xenopus laevis as a Model to Identify Translation Impairment

Published on: September 27, 2015

Various effects of paromomycin on tmRNA-mediated trans-translation

Takayuki Konno1, Toshiharu Takahashi, Akira Muto

  • 1Department of Biochemistry and Biotechnology, Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki 036-8561, Japan.

Nucleic Acids Research. Supplement (2001)
|September 27, 2003
PubMed

Insights

Paromomycin antibiotic affects bacterial trans-translation by binding tmRNA, inhibiting its function. This binding alters translation resumption and differs from paromomycin's effect on standard protein synthesis.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Biochemistry

Background:

  • Trans-translation is a unique cellular mechanism essential for rescuing stalled ribosomes.
  • tmRNA (transfer-messenger RNA) acts as both tRNA and mRNA, facilitating the release of stalled polypeptide chains.
  • Aminoglycoside antibiotics like paromomycin are known to interfere with bacterial translation.

Purpose of the Study:

  • To investigate the in vitro effects of paromomycin on tmRNA-mediated trans-translation.
  • To identify specific binding sites of paromomycin on tmRNA.
  • To elucidate the impact of paromomycin binding on key trans-translation events.

Main Methods:

  • Chemical footprinting to map paromomycin binding sites on tmRNA.
  • In vitro assays to assess the effects of paromomycin on tmRNA aminoacylation.
  • Analysis of translation resumption point shifts induced by paromomycin.
  • Site-directed mutagenesis of 16S rRNA and tmRNA to identify resistance mutations.

Main Results:

  • Paromomycin binds to tmRNA in both the tRNA domain and the helix connecting tRNA and mRNA domains.
  • Binding in the tRNA domain inhibits aminoacylation, an effect partially rescued by SmpB protein.
  • Paromomycin induces a -1 shift in the translation resumption point on tmRNA.
  • This shift is dependent on the 16S rRNA binding site but not tmRNA sequence variations.

Conclusions:

  • Paromomycin directly interferes with tmRNA function through specific binding.
  • The antibiotic's mechanism against trans-translation is distinct from its action on canonical translation.
  • Understanding these interactions could inform the development of new antibiotics targeting bacterial protein synthesis.

More Related Videos

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses11:19

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Published on: February 25, 2011

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling10:00

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Related Concept Videos

Translation01:37

Translation

Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.Translation Produces the Building Blocks of LifeProteins are called the...
Translation01:37

Translation

Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.Translation Produces the Building Blocks of LifeProteins are called the...
Leaky Scanning02:28

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Initiation of Translation02:33

Initiation of Translation

Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...