Related Experiment Video
Updated: Jun 12, 2025
09:04
Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
Published on: September 21, 2017
9.5K
Stapled Peptides as Inhibitors of mRNA Deadenylation
Sunit Pal1, Ilja Gordijenko1, Stefan Schmeing1
1Chemical Genomics Centre, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227, Dortmund, Germany.
Angewandte Chemie (International Ed. in English)
|September 25, 2024
Summary
Researchers developed a novel peptide, NIP-2, to inhibit messenger RNA (mRNA) degradation by targeting the CCR4-NOT complex. This approach stabilizes beneficial mRNA, offering a new therapeutic strategy to increase mRNA levels.
Area of Science:
- Molecular Biology
- Biochemistry
- Drug Discovery
Background:
- Therapeutic strategies often focus on reducing disease-causing mRNA.
- Stabilizing beneficial mRNA by preventing its degradation is an underexplored area.
- mRNA degradation, initiated by poly(A) tail removal, is regulated by the CCR4-NOT complex.
Purpose of the Study:
- To explore methods for blocking the destruction of beneficial mRNA.
- To develop a peptide-based inhibitor targeting the CCR4-NOT complex's deadenylation activity.
- To investigate the potential of stabilizing mRNA for therapeutic benefit.
Main Methods:
- Identified the NOT9 binding motif in the CCR4-NOT complex as a target.
- Designed and synthesized hydrocarbon stapled peptides, starting with NIP-2.
- Utilized co-crystallography for peptide optimization and in vitro/cell-based assays to assess inhibition and cell permeability.
Main Results:
- Developed NIP-2 peptide with nanomolar affinity for NOT9 and potent inhibition of RNA binding and deadenylation in vitro.
- Demonstrated cell permeability of NIP-2 (EC50 = 2.44 μM).
- Optimized peptide NIP-2-H27A-N3 showed improved affinity and significantly enhanced cell permeability (EC50 = 0.34 μM), inhibiting mRNA deadenylation in HeLa cells.
Conclusions:
- Peptide-based inhibition of the CCR4-NOT complex is feasible.
- The developed peptides can effectively inhibit mRNA deadenylation and increase mRNA stability.
- This strategy presents a novel therapeutic avenue for conditions requiring increased levels of specific beneficial mRNAs.
Related Concept Videos
02:51mRNA Stability and Gene Expression
5.6K
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
Cis-acting Elements involved in mRNA stability
5.6K
02:27Nonsense-mediated mRNA Decay
10.6K
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,...
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
10.6K
02:31Nuclear Export of mRNA
7.6K
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...
7.6K
01:53RNA Stability
33.4K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
33.4K
02:15Experimental RNAi
6.1K
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
6.1K
02:24Regulation of Expression Occurs at Multiple Steps
22.5K
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.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
22.5K