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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 at Multiple Steps01:23

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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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Regulation of Expression Occurs at Multiple Steps02:24

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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.
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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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Master Transcription Regulators02:23

Master Transcription Regulators

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Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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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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Describing a Transcription Factor Dependent Regulation of the MicroRNA Transcriptome
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TENT2, TUT4, and TUT7 selectively regulate miRNA sequence and abundance.

Acong Yang1, Xavier Bofill-De Ros1, Ryan Stanton1

  • 1RNA Mediated Gene Regulation Section; RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD, 21702, USA.

Nature Communications
|September 7, 2022
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Tailor-Nuclease (TENT) enzymes create specific microRNA (miRNA) isoforms through 3' tailing. This study reveals TENT2, TUT4, and TUT7 enzymes selectively modify miRNAs, impacting their function and regulation.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Terminal uridylate/adenylyl transferases (TENTs) are known to generate microRNA (miRNA) isoforms via 3' tailing.
  • The precise mechanisms and functional consequences of TENT-mediated miRNA tailing remain largely uncharacterized.

Purpose of the Study:

  • To elucidate the specific roles of TENT2, TUT4, and TUT7 in miRNA 3' tailing and subsequent functional regulation.
  • To investigate the impact of different tailing modifications (adenylation, uridylation, guanylation) on miRNA stability and activity.

Main Methods:

  • Generation of isogenic HEK293T cell lines with individual or combined knockouts of TENT2, TUT4, and TUT7.
  • Deep sequencing, Northern blot analysis, and in vitro assays to characterize TENT-specific effects on miRNA isoforms.
  • Rescue experiments to validate findings from knockout studies.

Main Results:

  • 3' tailing of miRNAs is a highly specific process, not random.
  • TENT2 exhibits adenylation, guanylation, and uridylation activity on mature miRNAs.
  • TUT4 is responsible for the uridylation of most miRNAs, while TUT7 appears dispensable for this process.
  • Abolishing uridylation significantly dysregulates specific miRNAs, whereas removing adenylation has a minor effect.
  • Distinct regulatory mechanisms by TUT4/7 impact miR-181b and miR-222, while TUT7 specifically upregulates the miR-888 cluster.

Conclusions:

  • TENT enzymes play selective roles in generating 3' miRNA isomiRs.
  • Uridylation is a critical modification for the regulation of a subset of miRNAs.
  • These findings provide a foundation for exploring the functional significance of TENT-mediated miRNA modifications.