The main aims of the project are: 1) to identify novel yeast proteins that are involved directly or indirectly in the mechanisms or regulation of various aspects of the production and processing of nuclear RNA, including transcription, 5' capping, splicing, 3' end processing/ polyadenylation and transport between the nucleus and the cytoplasm through the nucleopore, 2) to determine patterns of interaction between previously characterised and newly identified proteins, 3) to use these data to assign tentative functions to the newly identified ORFs, 4) to detect and investigate functional links between different pathways of RNA metabolism, for example between transcription and splicing, or between splicing and export, 5) to detect links between these pathways and other cellular processes, for example between the cell cycle and splicing or transcription, and to identify the factors making the connection(s), 6) to improve and optimise the technology of the existing two-hybrid method, 7) to evaluate the efficacy of the two-hybrid approach for obtaining these kinds of data, 8) to develop a novel twohybrid method based on activation of RNA polymerase III that will permit this approach to be extended to the analysis of polymerase II transcriptlon factors, 9) to produce computer programs that will facilitate the collection and analysis of sequence data from the fusion proteins, and 10) to assess the feasibility of extending this protein linkage analysis to other systems, either to other biochemical pathways in yeast or to higher eukaryotes.
The ultimate aim is to produce a detailed linkage map representing physical interactions between proteins that are functionally related, having roles in the mechanisms or regulation of pathways involved in RNA metabolism. The partners in this project are experts in complementary areas of this field: transcription, RNA splicing, 3' end processing and polyadenylation, nuclearcytoplasmic transport through the nucleopore structure. Each of these cellular processes is extremely complex, involving interactions between many factors. Only a minority of the genes encoding these factors have been identified to date and we expect to find many more in the course of this project. Therefore, we will propose functions for uncharacterised ORFs found in the yeast genome sequencing project.
Funding SchemeCSC - Cost-sharing contracts
91198 Gif Sur Yvette
EH9 3Jr Edinburgh