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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

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Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
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Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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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...
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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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siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
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Translational Regulation01:29

Translational Regulation

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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A Slow Dynamic RNA Switch Regulates Processing of microRNA-21.

Matthew D Shortridge1, Greg L Olsen1, Wen Yang2

  • 1Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA.

Journal of Molecular Biology
|June 25, 2022
PubMed
Summary
This summary is machine-generated.

Researchers discovered a slow conformational change in pre-microRNA-21 that affects its processing. This finding reveals a new RNA regulation mechanism and suggests chemical methods to control microRNA activity in disease.

Keywords:
NMRdynamicsmiRNA biogenesispre-miR-21regulation

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biochemistry

Background:

  • MicroRNAs (non-coding RNAs) regulate gene expression post-transcriptionally.
  • Dysregulation of microRNAs is implicated in various human diseases.
  • Understanding microRNA processing is crucial for therapeutic development.

Purpose of the Study:

  • To investigate the dynamic conformational changes at the Dicer cleavage site of pre-miR-21.
  • To identify regulatory mechanisms influencing pre-miR-21 processing.
  • To explore potential small molecule or peptide-based modulation of microRNA processing.

Main Methods:

  • Characterization of pre-miR-21 conformational dynamics using biophysical techniques.
  • Analysis of the impact of single nucleotide mutations on RNA structure and processing.
  • Screening of small molecule and peptide libraries for ligands binding to pre-miR-21.
  • Assays to measure microRNA processing efficiency in the presence of identified ligands.

Main Results:

  • Discovery of a slow conformational rearrangement (0.1 s⁻¹) at the Dicer cleavage site of pre-miR-21.
  • This dynamic switch regulates the equilibrium between readily and inefficiently processed RNA structural states.
  • Single nucleotide mutations were shown to alter this dynamic switch.
  • Small molecule and peptide ligands were identified that bias pre-miR-21 towards the inefficiently processed state, reducing processing efficiency.

Conclusions:

  • A novel mechanism of RNA regulation involving a slow conformational dynamics has been identified for pre-miR-21.
  • This dynamic switch provides a new target for modulating microRNA processing.
  • Chemical approaches using small molecules or peptides can be developed to suppress or activate pathogenic microRNAs by stabilizing specific conformational states.