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Related Concept Videos

Nucleotide Excision Repair01:38

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Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
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Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
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Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.
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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
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Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

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Untargeted Mutation Triggered by Ribonucleoside Embedded in DNA.

Tetsuya Suzuki1, Kiyoharu Yasui1, Yasuo Komatsu2

  • 1Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.

International Journal of Molecular Sciences
|January 8, 2025
PubMed
Summary

Incorporated ribonucleosides in DNA trigger mutations at guanine bases via an APOBEC3B-dependent pathway. This finding suggests a role for ribonucleosides in APOBEC3-dependent cancer initiation.

Keywords:
APOBEC3action-at-a-distance mutationribonucleoside

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA polymerases can misincorporate ribonucleoside 5'-triphosphates into DNA strands.
  • Incorporated ribonucleosides can lead to DNA damage and mutations.

Purpose of the Study:

  • To investigate the impact of incorporated ribonucleosides on untargeted mutations in human cells.
  • To elucidate the mechanism by which ribonucleosides induce mutations.

Main Methods:

  • Introduction of riboguanosine (rG) into the supF gene in a plasmid.
  • Transfection into U2OS cells and DNA recovery after 48 hours.
  • Mutation analysis using Escherichia coli RF01 strain and RNA interference for APOBEC3B knockdown.

Main Results:

  • Frequent untargeted base substitutions were observed at G bases within 5'-GpA-3' dinucleotides.
  • These mutations were reduced by approximately 80% upon APOBEC3B knockdown.
  • The mutation pattern resembled action-at-a-distance mutations induced by oxidative damage or APOBEC3 activity.

Conclusions:

  • Embedded ribonucleosides induce base substitution mutations at G bases through an APOBEC3B-dependent mechanism.
  • Ribonucleosides may contribute to APOBEC3-dependent cancer initiation events.