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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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Transfer RNA Synthesis02:36

Transfer RNA Synthesis

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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
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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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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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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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Structural Insights into the Human Pre-mRNA 3'-End Processing Machinery.

Yixiao Zhang1, Yadong Sun2, Yongsheng Shi3

  • 1Laboratory of Molecular Electron Microscopy, Rockefeller University, New York, NY 10065, USA.

Molecular Cell
|December 8, 2019
PubMed
Summary

The study reveals the core structure of mammalian pre-mRNA 3'-end processing machinery. Researchers identified a key protein interaction motif (PIM) essential for forming the cleavage and polyadenylation specificity factor (CPSF).

Keywords:
CPSF160CPSF73alternative polyadenyltationcleavage and polyadenylationpolyadenylation signal

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Area of Science:

  • Molecular biology
  • Biochemistry
  • Structural biology

Background:

  • Mammalian pre-mRNA 3'-end processing is vital for gene expression.
  • The precise architecture of the involved protein machinery, including cleavage and polyadenylation specificity factor (CPSF) and cleavage stimulation factor (CstF), remains largely unknown.
  • CPSF comprises two distinct modules: mCF and mPSF.

Purpose of the Study:

  • To elucidate the architectural organization of the mammalian pre-mRNA 3'-end processing machinery.
  • To identify key interactions within the CPSF complex.

Main Methods:

  • Production of recombinant human CPSF and CstF.
  • Electron microscopy (EM) and cryo-electron microscopy (cryo-EM) for structural analysis.
  • Site-directed mutagenesis to investigate protein interactions.

Main Results:

  • The polyadenylation specificity factor (mPSF) serves as the central organizing core of the machinery.
  • The conformations of the cleavage factor (mCF) and CstF are highly variable relative to mPSF.
  • A novel PSF interaction motif (PIM) within CPSF100 was identified, crucial for tethering mCF to mPSF.
  • Mutations in the PIM disrupt CPSF formation, confirming its critical role.

Conclusions:

  • The mPSF module is the structural anchor of the pre-mRNA 3'-end processing machinery.
  • The PIM is essential for the assembly and integrity of the CPSF complex.
  • Understanding this machinery's architecture provides insights into gene regulation.