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Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

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Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
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The Unfolded Protein Response01:37

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The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
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RNA Splicing

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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Riboswitches01:56

Riboswitches

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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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Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
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Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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Related Experiment Video

Updated: Jul 4, 2025

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

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Xrp1 governs the stress response program to spliceosome dysfunction.

Dimitrije Stanković1, Luke S Tain1, Mirka Uhlirova1

  • 1Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne 50931, Germany.

Nucleic Acids Research
|February 2, 2024
PubMed
Summary

Deficiency in U5 small nuclear ribonucleoprotein particles (snRNPs) causes R-loops and cell cycle arrest. The Xrp1-Irbp18 heterodimer mediates this stress response, highlighting splicing

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ACT1-CUP1 Assays Determine the Substrate-Specific Sensitivities of Spliceosomal Mutants in Budding Yeast
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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Co-transcriptional pre-mRNA processing by the spliceosome is crucial for gene expression and genome stability.
  • U5 small nuclear ribonucleoprotein particles (snRNPs) are essential components of the spliceosome.

Purpose of the Study:

  • To investigate the consequences of U5 snRNP deficiency in Drosophila imaginal cells.
  • To identify the molecular mechanisms linking spliceosome malfunction to cellular stress responses.

Main Methods:

  • Generating U5 snRNP-deficient Drosophila imaginal cells.
  • Transcriptome analysis (RNA-seq).
  • R-loop detection and quantification.
  • Western blotting for stress markers (JNK, p53).
  • RNA interference (RNAi) to knockdown Xrp1 and Irbp18.

Main Results:

  • U5 snRNP deficiency led to widespread transcriptome changes and mutagenic R-loop accumulation.
  • Cells exhibited stress responses, cell cycle arrest, increased protein translation, cell size, and apoptosis.
  • The Xrp1-Irbp18 heterodimer was identified as a key mediator of the stress program.
  • Knockdown of Xrp1 or Irbp18 reduced stress signaling, restored cell cycle, and inhibited apoptosis, but did not fix splicing defects.

Conclusions:

  • U5 snRNP malfunction triggers a DNA damage response pathway involving R-loops and the Xrp1-Irbp18 heterodimer.
  • Accurate splicing is essential for maintaining cellular and tissue homeostasis.
  • The Xrp1-Irbp18 heterodimer acts as a sensor for spliceosome defects and mediates stress-induced cellular senescence.