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Related Concept Videos

RNA Splicing01:32

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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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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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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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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.
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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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Using the E1A Minigene Tool to Study mRNA Splicing Changes
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Exon junction complex-associated multi-adapter RNPS1 nucleates splicing regulatory complexes to maintain

Lena P Schlautmann1,2, Jan-Wilm Lackmann3, Janine Altmüller4

  • 1Institute for Genetics, University of Cologne, 50674 Cologne, Germany.

Nucleic Acids Research
|May 31, 2022
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Summary

The exon junction complex (EJC)-binding protein RNPS1 primarily regulates splicing, not nonsense-mediated decay (NMD). RNPS1 ensures correct splicing of vulnerable events by forming diverse regulatory complexes.

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Gene Regulation

Background:

  • The exon junction complex (EJC) is crucial for post-transcriptional gene regulation, deposited during splicing.
  • RNPS1, an EJC-binding protein, has proposed roles in splicing and nonsense-mediated mRNA decay (NMD).

Purpose of the Study:

  • To investigate the primary function of RNPS1 in gene regulation.
  • To elucidate RNPS1's role in splicing and NMD through transcriptome-wide analysis and interactome mapping.

Main Methods:

  • Transcriptome-wide analysis of EJC and RNPS1 knockdown effects in human cell lines.
  • Complementary immunoprecipitations and proximity labeling to define the RNPS1 interactome.
  • Rescue experiments to assess the functional contribution of RNPS1 domains.

Main Results:

  • RNPS1 moderately influences NMD but is not essential for the process.
  • Aberrant splicing events upon RNPS1 knockdown indicate its primary role in splicing regulation.
  • RNPS1's RRM and C-terminal domains partially regulate splicing events.
  • Identified interactions with splicing factors dependent on specific RNPS1 domains.

Conclusions:

  • RNPS1 functions mainly as a splicing regulator, not a global NMD factor.
  • RNPS1 promotes accurate and efficient splicing of vulnerable sites through diverse complex formation.