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Transfer RNA Synthesis02:36

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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
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Nucleoside modification-based flexizymes with versatile activity for tRNA aminoacylation.

Xin-Dan Zhang1, Yi-Shen Wang1, Hua Xiang1

  • 1School of Pharmaceutical Sciences, South-Central Minzu University, Wuhan 430074, China. leixx@lzu.edu.cn.

Chemical Communications (Cambridge, England)
|January 17, 2024
PubMed
Summary

Researchers developed modified flexizymes (Fxs) using nucleoside modifications. These engineered ribozymes expand the genetic code by enabling tRNA charging with diverse substrates for synthetic biology.

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Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Molecular Biology

Background:

  • Flexizymes (Fxs) are ribozymes crucial for genetic code reprogramming via tRNA acylation.
  • Expanding the repertoire of amino acids incorporated into proteins is a key goal in synthetic biology.

Purpose of the Study:

  • To develop novel flexizyme variants with enhanced substrate acceptance.
  • To create a versatile platform for expanding the genetic code through modified tRNA acylation.

Main Methods:

  • A nucleoside-modification strategy was employed to synthesize flexizyme variants.
  • Key modifications included 2'-OMe, 2'-F, and 2'-MOE groups on the flexizyme structure.
  • The activity of these variants in charging tRNAs with diverse substrates was evaluated.

Main Results:

  • Novel flexizyme variants with unique and versatile activities were successfully prepared.
  • These modified flexizymes demonstrated the ability to charge tRNAs with a broad range of non-canonical substrates.
  • The modifications conferred enhanced catalytic properties for tRNA acylation.

Conclusions:

  • Nucleoside modification is an effective strategy for engineering flexizyme variants.
  • The developed flexizyme platform significantly expands the scope of substrates for genetic code expansion.
  • This work provides a robust method for incorporating diverse molecules into proteins, advancing synthetic biology.