Related Experiment Video
Updated: Jul 16, 2026
11:48
Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
Published on: October 9, 2014
RNA recognition and cleavage by a splicing endonuclease.
Song Xue1, Kate Calvin, Hong Li
1Department of Chemistry and Biochemistry, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA.
Science (New York, N.Y.)
|May 13, 2006
Summary
The RNA splicing endonuclease structure reveals how it cleaves precursor RNAs. This enzyme uses a cooperative mechanism, with its domains working together to prepare RNA for intron removal and protein synthesis.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- RNA splicing is crucial for producing functional RNAs.
- RNA splicing endonucleases are key enzymes in intron removal.
- Understanding endonuclease structure-function relationships is vital for deciphering gene expression regulation.
Purpose of the Study:
- To elucidate the structural basis of RNA splicing endonuclease activity.
- To characterize the interaction between the Archaeglobus fulgidus splicing endonuclease and its RNA substrate.
- To understand the mechanism of cooperative RNA cleavage.
Main Methods:
- X-ray crystallography was used to determine the structure of the endonuclease-RNA complex.
- High-resolution structural analysis (2.85 angstroms) was performed.
- Biochemical assays were implied to study the cleavage reaction.
Main Results:
- The structure of the Archaeglobus fulgidus splicing endonuclease bound to a bulge-helix-bulge RNA was determined.
- The endonuclease dimer was observed to recognize a flipped-out bulge base and stabilize bent RNA backbones.
- A cooperative mechanism involving an arginine pair from one catalytic domain sandwiching a nucleobase from the other domain was identified.
Conclusions:
- The determined structure provides atomic-level insights into the RNA splicing endonuclease mechanism.
- Cooperative recognition and stabilization of the RNA substrate are critical for efficient cleavage.
- This study advances our understanding of RNA processing and its regulation.
Related Concept Videos
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
03:21RNA-seq
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
01:32Pre-mRNA Processing: RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...