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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.
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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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pre-mRNA Processing

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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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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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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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Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
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PRMT5 promotes full-length HTT expression by repressing multiple proximal intronic polyadenylation sites.

Manisha Yadav1,2, Mona A AlQazzaz3, Felipe E Ciamponi4

  • 1Department of Medical Biophysics, University of Toronto, Toronto, ON, M5G1L7, Canada.

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Researchers identified PRMT5 as a novel regulator of Huntingtin (HTT) gene splicing. Inhibiting PRMT5 disrupts HTT mRNA processing, potentially lowering pathogenic protein levels and inducing neuronal differentiation.

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Area of Science:

  • Neuroscience
  • Genetics
  • Molecular Biology

Background:

  • Huntington's disease (HD) is caused by CAG repeat expansion in the Huntingtin (HTT) gene, leading to toxic polyglutamine-expanded HTT protein.
  • Current therapeutic strategies for HD focus on reducing HTT protein levels, with splice modulators showing promise.
  • Understanding HTT gene regulation is crucial for developing effective HD treatments.

Purpose of the Study:

  • To identify novel regulators of HTT mRNA splicing and alternative polyadenylation.
  • To investigate the role of PRMT5 in HTT gene expression.
  • To explore the therapeutic potential of targeting PRMT5 for Huntington's disease.

Main Methods:

  • Investigated PRMT5 as a regulator of HTT splicing and alternative polyadenylation.
  • Analyzed the impact of PRMT5 inhibition on HTT mRNA processing, including intron splicing and polyadenylation.
  • Examined HTT transcript levels during neuronal differentiation and in response to PRMT5 inhibition in glioblastoma stem cells.

Main Results:

  • PRMT5 was identified as a novel regulator of HTT mRNA splicing and alternative polyadenylation.
  • PRMT5 inhibition disrupted HTT intron 9 and 10 splicing, activating intronic polyadenylation sites and promoting premature mRNA termination.
  • Truncated HTT transcripts increased during neuronal differentiation, correlating with decreased PRMT5 expression; PRMT5 inhibition induced neuronal differentiation in glioblastoma stem cells.

Conclusions:

  • PRMT5 plays a significant role in modulating HTT mRNA expression through regulation of intronic polyadenylation and premature termination.
  • PRMT5 inhibition offers a potential therapeutic strategy for Huntington's disease by reducing pathogenic HTT protein levels.
  • PRMT5-mediated regulation of HTT mRNA is important during neuronal differentiation.