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Related Concept Videos

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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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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mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
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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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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.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
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Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Related Experiment Video

Updated: Oct 18, 2025

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
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Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

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Dynamics in Fip1 regulate eukaryotic mRNA 3' end processing.

Ananthanarayanan Kumar1, Conny W H Yu1, Juan B Rodríguez-Molina1

  • 1MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.

Genes & Development
|October 1, 2021
PubMed
Summary
This summary is machine-generated.

The study reveals how yeast Fip1 anchors poly(A) polymerase Pap1 to the cleavage and polyadenylation factor (CPF) complex. Fip1

Keywords:
CPFCPSFRNA-binding proteindynamicsmRNA processingpolyadenylation

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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing RIPiT-Seq
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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • The cleavage and polyadenylation factor (CPF) complex is crucial for mRNA 3' end processing in eukaryotes.
  • CPF comprises multiple subunits, some of which possess intrinsically disordered regions (IDRs).
  • Intrinsically disordered regions can exhibit flexibility or adopt ordered structures upon binding partners.

Purpose of the Study:

  • To investigate the role of intrinsically disordered regions in CPF function.
  • To elucidate the interaction between Fip1, Pap1, and Yth1 within the CPF complex.
  • To determine the dynamics of Fip1 within the CPF complex during mRNA processing.

Main Methods:

  • Reconstitution of a recombinant 850-kDa CPF complex.
  • Selective labeling of Fip1 for incorporation into the recombinant CPF.
  • Nuclear magnetic resonance (NMR) spectroscopy to study Fip1 dynamics within CPF.

Main Results:

  • Yeast Fip1 anchors the poly(A) polymerase Pap1 to CPF through an interaction with Yth1's zinc finger 4.
  • A Fip1 intrinsically disordered region connecting Yth1- and Pap1-binding sites remains highly dynamic within the CPF complex.
  • The dynamics of Fip1 are essential for coordinating cleavage and polyadenylation.

Conclusions:

  • Fip1's dynamic intrinsically disordered region is key to its function in CPF.
  • The flexibility of Fip1 facilitates the coordination of mRNA cleavage and polyadenylation.
  • Understanding Fip1 dynamics provides insights into the regulation of mRNA 3' end processing.