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Related Concept Videos

From DNA to Protein03:06

From DNA to Protein

The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
Transfer RNA Synthesis02:36

Transfer RNA Synthesis

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.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
tRNA Activation02:26

tRNA Activation

Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
Improving Translational Accuracy02:07

Improving Translational Accuracy

Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
Transfer RNA Synthesis02:36

Transfer RNA Synthesis

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.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
tRNA Activation02:26

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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...

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Related Experiment Video

Updated: Jul 21, 2026

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
12:36

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Published on: October 19, 2010

Designer ribozymes: programming the tRNA specificity into flexizyme.

Krishna Ramaswamy1, Hirohide Saito, Hiroshi Murakami

  • 1Departments of Chemistry and Biological Sciences, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA.

Journal of the American Chemical Society
|September 16, 2004
PubMed
Summary

Researchers engineered a new artificial enzyme, Fx10, to specifically attach amino acids to transfer RNAs (tRNAs). This custom catalyst enables precise creation of nonnatural aminoacyl-tRNAs for advanced biotechnology.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Synthetic Biology

Background:

  • Artificial ribozymes like Fx3 can aminoacylate transfer RNAs (tRNAs) with phenylalanine and its derivatives.
  • Generic ribozymes lack specificity, limiting their application in precise synthesis.

Purpose of the Study:

  • To engineer a generic ribozyme (Fx3) with enhanced tRNA specificity.
  • To create a customizable catalytic system for generating nonnatural aminoacyl-tRNAs.

Main Methods:

  • Appending a tRNA-specific sequence (TSS) to the 3'-end of the Fx3 ribozyme.
  • Designing the TSS to be complementary to the acceptor stem of a target tRNA.
  • Characterizing the interaction between the modified ribozyme (Fx10) and cognate/noncognate tRNAs.

Main Results:

  • The new designer ribozyme, Fx10, specifically recognizes and aminoacylates its cognate tRNA.
  • Fx10 utilizes a 10-base-pair interaction formed by TSS invasion of the tRNA acceptor stem.
  • Fx10 demonstrates high specificity, effectively discriminating against noncognate tRNAs.

Conclusions:

  • A simple strategy was developed to impart tRNA specificity to artificial ribozymes.
  • Fx10 serves as a programmable, custom-made catalyst for producing specific nonnatural aminoacyl-tRNAs.
  • This approach facilitates the generation of diverse nonnatural aminoacyl-tRNAs for biotechnological applications.