Splicing

The process of gene expression in the cell is carried out by a series of "protein machines." The process starts with DNA transcription (reading the "blueprint" for a protein on a particular gene) and carries through to production of that protein by ribosomes in the cytoplasm.

The stepwise production of a protein begins with an RNA copy of the DNA blueprint. This RNA cannot be translated into protein by the cells until certain stretches of information which are not required to make the protein are cut or "spliced" out of it.

Splicing is performed by a protein machine termed the "spliceosome", a complex of more than 50 proteins acting together to catalyse this multi-step reaction. One of our main research goals is to shed further light on the organisation of this structure and its regulation within the cell nucleus. Until we understand how it functions normally, we cannot understand what has gone wrong in disease states which affect this particular cellular process.



For most eukaryotic genes that code for proteins, splicing of precursor transcripts (pre-mRNA) is required to generate functional mRNA products. During splicing, the pre-mRNA substrate is assembled into a large complex, termed a spliceosome, whose major subunits are small nuclear ribonuclear protein particles (snRNPs); in particular U1, U2, U4/6 and U5 snRNPs. The snRNPs act in conjunction with additional,non-snRNP protein splicing factors to catalyse excision of introns from pre-mRNA.

We have purified mammalian spliceosomes and analysed their protein composition by 2-D gel electrophoresis and electrospray mass spectrometry. This analysis has greatly facilitated functional studies on the splicing mechanism. We are also analysing how reversible protein phosphorylation can regulate the assembly of spliceosomes and the catalysis of splicing.

For more information about in vitro assays used to measure splicing activity, visit our Protocols page.

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