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Related Concept Videos

pre-mRNA Processing02:01

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.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
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Chromatin Structure Regulates pre-mRNA Processing02:41

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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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Pre-mRNA Processing: Modification of pre-mRNA Ends01:35

Pre-mRNA Processing: Modification of pre-mRNA Ends

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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.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
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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...
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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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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

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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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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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Bulges control pri-miRNA processing in a position and strand-dependent manner.

Shaohua Li1, Thi Nhu-Y Le1, Trung Duc Nguyen1

  • 1Division of Life Science, The Hong Kong University of Science & Technology, Hong Kong, China.

RNA Biology
|December 31, 2020
PubMed
Summary

Bulges in primary microRNA (pri-miRNA) structures are crucial for accurate processing by the Microprocessor complex. Specific bulges, like midB, enhance microRNA (miRNA) production by orienting DROSHA, improving gene expression regulation.

Keywords:
BulgesDGCR8DROSHAMicroprocessormiRNA biogenesis

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) regulate gene expression and are implicated in various human diseases.
  • The biogenesis of miRNAs depends on the processing of primary miRNA (pri-miRNA) transcripts by the Microprocessor complex, comprising DROSHA and DGCR8.
  • Understanding pri-miRNA processing is key to deciphering miRNA biogenesis and its regulatory roles.

Purpose of the Study:

  • To investigate the role of bulges within pri-miRNA secondary structures in the processing activity of the Microprocessor complex.
  • To determine how bulges influence the efficiency and accuracy of pri-miRNA cleavage.
  • To elucidate the mechanism by which specific bulges, such as midB, affect DROSHA orientation and subsequent miRNA production.

Main Methods:

  • Analysis of high-throughput pri-miRNA processing assays.
  • Examination of pri-miRNA secondary structures.
  • Investigating the positional and strand-dependent effects of bulges on Microprocessor activity.
  • Characterizing the function of conserved bulges (midB) in DROSHA orientation.

Main Results:

  • Bulges at various positions within pri-miRNAs significantly control both the efficiency and accuracy of Microprocessor-mediated cleavage.
  • These bulges exert their effects on the Microprocessor complex, specifically interacting with the catalytic subunit DROSHA.
  • The function of bulges is dependent on their position and the strand of the pri-miRNA.
  • Enriched and conserved bulges, termed midB, were found to correct DROSHA orientation on pri-miRNAs, leading to enhanced miRNA production.

Conclusions:

  • Bulges within pri-miRNA structures are critical regulatory elements in miRNA biogenesis.
  • The position and sequence context of bulges dictate their impact on DROSHA-DGCR8 processing.
  • The midB bulge represents a novel mechanism for enhancing miRNA production by optimizing DROSHA engagement.
  • These findings provide a deeper understanding of pri-miRNA processing and highlight potential targets for miRNA biogenesis regulation.