| 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. |