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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...
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...
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.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...

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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

Splicing factors SF1 and U2AF associate in extraspliceosomal complexes.

José Rino1, Joana M P Desterro, Teresa R Pacheco

  • 1Instituto de Medicina Molecular, Faculdade de Medicina, Av. Prof. Egas Moniz, 1649-028 Lisboa, Portugal.

Molecular and Cellular Biology
|February 21, 2008
PubMed
Summary

Splicing factors SF1 and U2AF form stable complexes, influencing spliceosome assembly. These interactions occur independently of splicing, suggesting a broader role in RNA processing beyond catalytic spliceosomes.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Splicing factors SF1 and U2AF are essential for recognizing the 3' splice site during spliceosome assembly.
  • SF1 is displaced during spliceosome remodeling, stabilizing U2 snRNP binding to pre-mRNA.

Purpose of the Study:

  • To investigate the interaction dynamics and cellular localization of splicing factors SF1 and U2AF.
  • To determine if the SF1-U2AF interaction is dependent on active splicing.

Main Methods:

  • Fluorescence microscopy to observe protein distribution in live cells.
  • Fluorescence recovery after photobleaching (FRAP) to analyze protein mobility.
  • Fluorescence resonance energy transfer (FRET) to detect direct protein binding.

Main Results:

  • SF1 and U2AF are found in the nucleoplasm and nuclear speckles.
  • Protein mobility of SF1 and U2AF subunits correlates with their interaction.
  • Direct binding between SF1 and U2AF(65) was confirmed via FRET.
  • This interaction persists even after transcription inhibition.

Conclusions:

  • SF1 and U2AF form stable, splicing-independent complexes.
  • These extraspliceosomal complexes may play roles before and after participation in catalytic spliceosomes.
  • The findings provide insights into the dynamic assembly and regulation of the spliceosome.