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Related Concept Videos

RNA Splicing01:32

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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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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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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Using the E1A Minigene Tool to Study mRNA Splicing Changes
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Transcriptome-wide modulation of splicing by the exon junction complex.

Zhen Wang, Valentine Murigneux, Hervé Le Hir

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    |December 6, 2014
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    Summary
    This summary is machine-generated.

    The exon junction complex (EJC) core proteins regulate widespread alternative splicing in mammalian cells. This regulation is linked to RNA polymerase II elongation rates, revealing a broader role for EJC in splicing.

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    Area of Science:

    • Molecular Biology
    • RNA Biology
    • Gene Expression Regulation

    Background:

    • The exon junction complex (EJC) is a protein complex deposited on spliced mRNAs.
    • Core EJC proteins (eIF4A3, Magoh, Y14, MLN51) bind mRNA throughout its lifecycle.
    • Previous studies suggested EJC involvement in specific splicing events.

    Purpose of the Study:

    • To investigate the role of EJC core proteins in global alternative splicing.
    • To determine if EJC core proteins directly regulate splicing patterns.
    • To explore the relationship between EJC and RNA polymerase II activity.

    Main Methods:

    • Knockdown of EJC core proteins using siRNA in mammalian cells.
    • Analysis of alternative splicing changes genome-wide.
    • Rescue experiments with siRNA-resistant eIF4A3.
    • Measurement of RNA polymerase II elongation rates.

    Main Results:

    • Knockdown of EJC core proteins induced widespread alternative splicing changes.
    • These changes were specific to EJC core proteins and distinct from other splicing factors.
    • Splicing alterations were rescued by siRNA-resistant eIF4A3.
    • Splicing changes correlated with altered RNA polymerase II elongation rates.

    Conclusions:

    • EJC core proteins play a broader role in regulating alternative splicing than previously known.
    • A link exists between messenger ribonucleoprotein (mRNP) assembly and splice site recognition.
    • EJC's function extends to global control of alternative splicing through modulation of transcription elongation.