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Related Concept Videos

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RNA Polymerase II Accessory Proteins

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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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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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RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
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Transcriptional Regulation: Riboswitches01:23

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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
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Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
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Related Experiment Video

Updated: Jun 6, 2025

In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection
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Influenza A virus transcription generates capped cRNAs that activate RIG-I.

Elizaveta Elshina1,2, Emmanuelle Pitre1,2, Marisa Mendes3

  • 1Department of Molecular Biology, Princeton University, Washington Road, Princeton, NJ 08544, USA.

Biorxiv : the Preprint Server for Biology
|November 28, 2024
PubMed
Summary

Influenza A virus (IAV) transcription generates a novel capped complementary RNA (ccRNA). This ccRNA, upon hybridizing with viral RNA, activates the retinoic acid-inducible gene I (RIG-I) pathway, enhancing innate immune responses.

Keywords:
RIG-IRNA polymeraseaberrant RNAcapped cRNAinfluenza A virustranscription

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Area of Science:

  • Virology
  • Immunology
  • Molecular Biology

Background:

  • Host pathogen receptor retinoic acid-inducible gene I (RIG-I) detects viral RNA to trigger innate immunity.
  • Innate immune responses can be initiated by viral transcription, but the mechanisms are not fully understood.

Purpose of the Study:

  • To investigate if immunostimulatory RNA is produced during influenza A virus (IAV) transcription.
  • To identify novel RNA species generated during IAV infection and their role in immune activation.

Main Methods:

  • Investigated IAV RNA polymerase activity during transcription termination.
  • Utilized in vitro assays, ribonucleoprotein reconstitution, and IAV infection models.
  • Analyzed RNA structures using molecular biology techniques and assessed RIG-I activation.

Main Results:

  • Discovered a novel capped complementary RNA (ccRNA) produced by IAV RNA polymerase read-through transcription.
  • ccRNA formation is enhanced by mutations disrupting polyadenylation signals, leading to increased RIG-I activation.
  • ccRNA acts as a RIG-I agonist when hybridized with complementary negative-sense viral RNA.

Conclusions:

  • Identified a novel non-canonical IAV RNA species, ccRNA.
  • Proposed an alternative mechanism for RIG-I activation during IAV infection involving ccRNA-mediated hybridization.