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Related Concept Videos

Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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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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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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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RNA Interference

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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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SmD3 regulates intronic noncoding RNA biogenesis.

Benjamin S Scruggs1, Carlos I Michel, Daniel S Ory

  • 1Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University, St. Louis, Missouri, USA.

Molecular and Cellular Biology
|August 8, 2012
PubMed
Summary

The spliceosomal protein SmD3 regulates small nucleolar RNA (snoRNA) expression, impacting cellular response to metabolic stress. This finding reveals a novel mechanism linking splicing factors to noncoding RNA processing and lipotoxicity.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Excess lipid accumulation in nonadipose tissues contributes to oxidative stress, organ dysfunction, and diabetic complications.
  • Previous studies identified small nucleolar RNAs (snoRNAs) as mediators of lipotoxic cell death under conditions of metabolic stress.

Purpose of the Study:

  • To identify novel molecular mechanisms underlying lipotoxicity and cellular response to metabolic stress.
  • To investigate the role of spliceosomal protein SmD3 in regulating noncoding RNA processing and its contribution to lipotoxicity.

Main Methods:

  • Retroviral promoter trap mutagenesis was employed to generate mutant Chinese hamster ovary (CHO) cell lines resistant to lipotoxic and oxidative stress.
  • Characterization of a novel SmD3 haploinsufficiency mutant to assess its impact on snoRNA expression and pre-mRNA splicing.

Main Results:

  • A novel mutant with disrupted SmD3 gene demonstrated reduced snoRNA expression and abundance of snoRNA-containing intron lariats.
  • SmD3 haploinsufficiency did not impede pre-mRNA splicing but affected small nuclear RNAs (snRNAs) U4 and U5 levels.
  • SmD3 was implicated as a critical regulator of intronic noncoding RNA processing and a mediator of metabolic stress response pathways.

Conclusions:

  • SmD3 plays a crucial role in the processing of intronic noncoding RNAs, particularly snoRNAs.
  • Regulation of snoRNA expression by SmD3 links splicing machinery to cellular metabolic stress responses.
  • This study identifies SmD3 as an upstream mediator in pathways combating lipotoxicity and associated organ dysfunction.