Angus I. Lamond
We are studying the functional organisation of the cell nucleus and the mechanism of pre-mRNA splicing in mammalian cells. The importance of understanding nuclear organisation is underlined by recent evidence showing that multiple human diseases, including inherited genetic disorders, malignancies and viral infections, modify or disrupt subnuclear bodies. Our aim is to understand how subnuclear structures assemble, how splicing factors and other proteins are targeted to them and how factors traffic in the nucleoplasm between separate structures.
In parallel, we are carrying out detailed functional studies on novel human nuclear proteins we have identified. The major subunits of spliceosomes are the RNA-protein complexes called snRNPs. In vivo, snRNPs show a complex and dynamic localisation pattern. We have shown that newly assembled splicing snRNPs accumulate in Cajal bodies prior to speckles when they are first imported into the nucleus. The data indicate a specific pathway whereby snRNPs interact with separate nuclear structures in a defined temporal sequence.
We are studying the dynamic behaviour of nuclear proteins and RNPs by expressing genes fused to fluorescent protein tags and performing time-lapse microscopy on living cells. We have developed protocols allowing the isolation of intact subnuclear bodies from human cells, including nucleoli and coiled bodies. We are using quantitative mass spectrometry and in vivo imaging techniques to characterise the protein components of these isolated nuclear bodies and we have developed methods using quantitative mass spectrometry to characterise specific protein interaction partners and to map the subcellular localisation of proteins in high throughput.
Key lab techniques: fluoresence imaging, FRET, FLIM, quantitative mass spectrometry, nuclear fractionation and pre-mRNA splicing assays.
Key lab reagents vectors expressing GFP-tagged nuclear proteins and stable HeLa cell lines expressing GFP-tagged proteins.
Lab contact: Ursula Ryder: email@example.com
Lab website: www.lamondlab.com