In this project, the investigator proposes to analyze the contribution of latent membrane protein 1 (LMP-1) of EBV to immortalization of cells. She will use molecular genetic and biochemical analyses to determine the transmembrane topology of LMP-1, to study the quaternary structure of LMP-1 and its relevance to function, and determine the relationship between the function of the amino -terminus of LMP-1 and its structure, cytoskeletal association, turnover, and membrane patching.
The specific aims of the research are (1) to determine the transmembrane topology of the LMP-1 protein in the immortalized cell, (2) to determine if homo- oligomerization is required for function of the LMP-1 in immortalization, and (3) to determine how the structure of the cytoplasmic N-terminus of LMP-1 relates to function and the biochemical properties of cytoskeletal association, membrane patching, and rapid turnover, all of which suggest receptor-ligand function. The investigator points out that while it is clear that the N-terminal of LMP-1 is responsible for transformation by EBV, there is controversy as to whether this is due simply to the structure of the molecule in the membrane, or due to protein-protein interactions. Eventually, the investigator hopes to determine how LMP-1 functions in signal transduction leading to immortalization of B cells.
