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Related Concept Videos

LTR Retrotransposons03:08

LTR Retrotransposons

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
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General Transcription Factors01:30

General Transcription Factors

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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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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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piRNA - Piwi-interacting RNAs02:57

piRNA - Piwi-interacting RNAs

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PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...
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Transcription Elongation Factors02:35

Transcription Elongation Factors

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Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
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Related Experiment Video

Updated: Sep 11, 2025

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

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ITAF45 is a pervasive trans-acting factor for picornavirus Type II IRES elements.

Michael A Bellucci1,2, Mehdi Amiri1,2, Stephen Berryman3

  • 1Department of Biochemistry, McGill University, Montreal H3A 1A3, QC, Canada.

Proceedings of the National Academy of Sciences of the United States of America
|August 11, 2025
PubMed
Summary
This summary is machine-generated.

The cellular protein ITAF45 is crucial for the replication of multiple picornaviruses, including FMDV and EMCV. Its removal via CRISPR/Cas9 knockout renders cells resistant to these viral infections, highlighting its potential as an antiviral target.

Keywords:
host factorpicornavirustranslation

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Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Picornaviruses, lacking a 5' cap, use internal ribosome entry sites (IRES) for translation.
  • IRES activity is modulated by host proteins known as IRES trans-acting factors (ITAFs).
  • ITAF45 was previously considered specific to foot-and-mouth disease virus (FMDV) infection.

Purpose of the Study:

  • To investigate the broader role of ITAF45 in Type II IRES-containing picornavirus replication.
  • To determine if ITAF45 is a general host factor for various picornaviruses.

Main Methods:

  • CRISPR/Cas9 gene editing to knockout ITAF45 in human cell lines.
  • Infection assays with FMDV, encephalomyocarditis virus (EMCV), Theiler's murine encephalomyelitis virus (TMEV), and equine rhinitis A virus (ERAV).
  • Analysis of ITAF45's role in enhancing translation initiation on Type II IRES elements.

Main Results:

  • CRISPR/Cas9 knockout of ITAF45 conferred resistance to FMDV, EMCV, TMEV, and ERAV infection.
  • ITAF45 was confirmed as a pervasive host factor for picornaviruses utilizing Type II IRES.
  • The C-terminal lysine-rich region of ITAF45 mediates viral RNA binding and enhances translation initiation.

Conclusions:

  • ITAF45 plays a significant, previously underestimated role in the replication of diverse Type II IRES picornaviruses.
  • These findings challenge prior assumptions about ITAF45's restricted function.
  • ITAF45 represents a promising pan-viral therapeutic target against animal and emerging human cardioviruses.