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Leaky Scanning02:28

Leaky Scanning

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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...
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The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
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Improving Translational Accuracy02:07

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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...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Exon Recombination02:32

Exon Recombination

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The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
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Related Experiment Video

Updated: Jul 6, 2025

Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics
09:23

Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics

Published on: January 5, 2024

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Predicting stop codon reassignment improves functional annotation of bacteriophages.

Ryan Cook1, Andrea Telatin1, George Bouras2,3

  • 1Food, Microbiome and Health Research Programme, Quadram Institute Bioscience, Norwich, NR4 7UQ, UK.

Biorxiv : the Preprint Server for Biology
|January 8, 2024
PubMed
Summary
This summary is machine-generated.

Many bacteriophages use alternate genetic codes, repurposing stop codons for amino acids. Our study developed tools to automatically predict this, significantly improving viral genome annotation and functional identification.

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Area of Science:

  • Virology
  • Genomics
  • Bioinformatics

Background:

  • Bacteriophage diversity is vast, with many uncharacterized genomes.
  • Some phage lineages, like Crassvirales, utilize alternate genetic codes by reassigning stop codons.

Approach:

  • We identified phage genomes repurposing stop codons in INPHARED and the Unified Human Gut Virome catalogue (UHGV).
  • Modified annotation tools, Pharokka-gv and Prokka-gv, were developed to predict stop codon reassignment before genomic annotation.
  • These tools were used to re-annotate affected viral sequences.

Key Points:

  • Pharokka-gv and Prokka-gv significantly improved annotation quality for phages using alternate genetic codes.
  • Re-annotation with Pharokka-gv increased median gene length by over 67% for UHGV and 72% for INPHARED sequences.
  • Coding density increased substantially, and the proportion of functionally annotated genes, including major capsid proteins, rose significantly.

Conclusions:

  • Automatic prediction of stop codon reassignment is crucial for accurate viral genome annotation.
  • These improved annotation methods benefit downstream viral genomic and metagenomic analyses.