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Related Concept Videos

Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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

Pre-mRNA Processing: Modification of pre-mRNA Ends

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 the cell...
pre-mRNA Processing02:01

pre-mRNA Processing

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 guanosine). This 5’ cap helps the...
Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

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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Related Experiment Video

Updated: Jun 16, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
09:16

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

Published on: May 3, 2014

Enzymatic processing of platinated RNAs.

Erich G Chapman1, Victoria J DeRose

  • 1Department of Chemistry, University of Oregon, Eugene, Oregon 97403, USA.

Journal of the American Chemical Society
|January 27, 2010
PubMed
Summary

The antitumor drug cisplatin forms platinum-RNA adducts, disrupting RNA processing enzymes like phosphodiesterases and reverse transcriptase. Thiourea can reverse these platinum-RNA adducts, offering a tool for future research.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Cisplatin is a widely used antitumor drug that primarily targets DNA.
  • The interaction of cisplatin with RNA and its impact on RNA processing enzymes are not well understood.
  • Understanding RNA-Pt adducts is crucial for comprehending the broader cellular effects of platinum-based chemotherapy.

Purpose of the Study:

  • To investigate how platinum-II (Pt(II)) adducts on RNA affect the activity of key RNA processing enzymes.
  • To determine the influence of [Pt(NH(3))(2)](2+)-RNA adducts on phosphodiesterases, endoribonucleases, and reverse transcriptase.
  • To explore the potential contribution of RNA platination to the cellular effects of cisplatin.

Main Methods:

  • Synthesis of RNA oligonucleotides with specific platinum adducts.

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Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
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Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

Published on: January 3, 2019

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events
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Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

Published on: May 13, 2019

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Last Updated: Jun 16, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
09:16

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

Published on: May 3, 2014

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
08:12

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

Published on: January 3, 2019

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events
10:59

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

Published on: May 13, 2019

  • Assays to measure the activity of 3'-->5' and 5'-->3' phosphodiesterases.
  • Enzymatic assays using a purine-specific endoribonuclease (RNase U2) and a reverse transcriptase.
  • Chemical reversal of platinum-RNA adducts using thiourea.

    • Main Results:

    • Single Pt(II) adducts on RNA oligonucleotides blocked exonucleolytic digestion.
    • Most endoribonucleolytic cleavages were disrupted by platinum adducts, with RNase U2 showing tolerance to platination at the GA site.
    • Platinum adducts on more complex RNA structures inhibited reverse transcription, indicating interference with RNA sequence information transfer.
    • Thiourea was demonstrated to effectively reverse cisplatin-RNA adducts.

    Conclusions:

    • Cisplatin-RNA adducts significantly impair the function of essential RNA processing enzymes.
    • Platinum-induced disruption of RNA processing may contribute to the overall cytotoxicity of cisplatin.
    • The findings highlight RNA as a potential target for cisplatin and suggest RNA processing alterations as a mechanism of action.
    • Thiourea provides a valuable chemical tool for further investigation of cisplatin-RNA interactions.