Research objectives and content
The aim of the project is the characterization of the factors involved in intron recognition and assembly of the spliceosome, a basic process of pre-mRNA processing. We want to understand the mechanisms by which the cell recognizes an intron and decides to cut it out. We want to assess the role of the different factors present in U1 snRNP, the first particle that binds the pre-mRNA. We will study the recently described U1 snRNP proteins by using biochemistry and molecular biology methods. We will use yeast as a model. The availability of its genome and the fact that it is widely used in research makes it suitable for our purposes. This project has been started funded with an EC institutional fellowship that covers only one year. From our preliminary results we have found a protein that is part of the U1 snRNP and seems to have an important function in pre-mRNA recognition. We have also started to study other 4 proteins present in U1 snRNP in order to address their role in this particle. I have already purified the U1 snRNP and we want to carry out biochemical reconstitution of this particle to get commitment complexes. This knowledge will allow us to understand how the splicing machinery is able to distinguish between fully spliced RNAs and intron-containing pre-mRNAs, a basic process in pre-mRNA maturation.
Training content (objective, benefit and expected impact)
During my Ph. D. Thesis I have acquired training in molecular biology techniques. This project will be a perfect complement in my formation since I will use molecular biology, biochemistry and yeast genetics methodology to address the questions mentioned above. The EMBL provides a good environment for research. It has any facility required to perform the research project. This together with its scientific atmosphere will contribute to my education in such a way that allows me to carry out independent research. The analysis of spliceosome assembly will allow us ultimately to understand how splicing catalysis and accurate splice site selection are achieved. This knowledge will give us clues about how alternative splicing is carried out in the cells. This knowledge has direct implications to understand the regulation of gene expression. Links with industry / industrial relevance (22)