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Related Concept Videos

RNA Splicing01:32

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...
RNA Splicing01:32

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...
Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Pre-mRNA Processing: RNA Splicing01:32

Pre-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...

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Related Experiment Video

Updated: Jul 14, 2026

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

Sequence requirements for splicing by the Cne PRP8 intein.

Esther J Pearl1, Joel D A Tyndall, Russell T M Poulter

  • 1Department of Biochemistry, University of Otago, Dunedin 9054, New Zealand.

FEBS Letters
|June 5, 2007
PubMed
Summary

Investigating conserved residues in the Cne PRP8 intein revealed essential roles for specific amino acids in protein splicing. A key finding shows W151 prevents premature cleavage, ensuring complete splicing.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Chemistry

Background:

  • Protein splicing is a post-translational modification where an internal protein segment (intein) excises itself and ligates flanking sequences (exteins).
  • The Cne PRP8 intein is a well-studied example, but the precise roles of its conserved residues in the splicing mechanism require further elucidation.

Purpose of the Study:

  • To determine the functional importance of conserved residues in the Cne PRP8 intein for protein splicing efficiency.
  • To identify novel residues critical for the catalytic activity and fidelity of the Cne PRP8 intein.

Main Methods:

  • Alanine scanning mutagenesis was employed to systematically replace conserved residues with alanine.
  • Mutant inteins were expressed within a foreign protein context to assess their splicing activity.
  • Comparative analysis of splicing efficiency and cleavage patterns among various mutants was performed.

Main Results:

  • Mutagenesis confirmed the essentiality of residues at splice junctions and conserved residues in motif B (threonine and histidine).
  • Five previously unrecognized residues were identified as absolutely indispensable for protein splicing.
  • The W151A mutant exhibited premature C-terminal cleavage, indicating a distinct role for this residue.

Conclusions:

  • Specific conserved residues, including T, H, and five newly identified ones, are critical for Cne PRP8 intein-mediated protein splicing.
  • Residue W151 plays a crucial role in preventing premature C-terminal cleavage, thereby promoting complete intein splicing over sequential cleavage events.
  • A model is proposed where W151 acts as a gatekeeper, ensuring the fidelity of the splicing process.