| Co-Investigator(Kenkyū-buntansha) |
NAGATA Satoshi Grad. Sch. Agriculture and Life Science, Univ. of Tokyo, Lecturer, 大学院・農学生命科学研究科, 講師 (40246682)
IHARA Sayoko Grad. Sch. Agriculture and Life Science, Univ. of Tokyo, Pfor., 大学院・農学生命科学研究科, 助手 (80292788)
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| Research Abstract |
Phosphatodylinosito1 3-kinase is the enzyme that produces PIPィイD23ィエD2 in vivo. PIP3ィイD2aィエD2 acts as an important second messenger to activate various downstream factors. However, despite the efforts of a number of laboratories, there are not enough downstream factors identified to explain variety of cell responses dependent of PI 3-kinase pathway. We have produced PIPィイD23ィエD2 analogue beads and used them for identification of PIPィイD23ィエD2 binding proteins, which are the candidates for the downstream factors of PIPィイD23ィエD2 PIP3BP is one of such PIPィイD23ィエD2 binding proteins which localizes in the nucleus. Since it is homologous to Arf-GAP, it is likely that this protein is involved in vesicle transport. This year, we established hybridoma lines which produce BP-kinesin, a binding protein for PIP3B. We also identified SWAP70 as a PIPィイD23ィエD2 binding protein which is suggested to be a subunit of a B-cell specific recombinase complex. SWAP70 localized in the cytoplasm but moved into nucleus when co-expressed with a constitutively active PI 3-kinase. This translocation was sensitive to wortmannin, PI 3-kinase inhibitor. SWAP70 was overexpressed in Sf9 cells and purified. In vitro analysis for nuclear translocation revealed that SWAP70 is impo1-ted to the nucleus in an ATP dependent manner. It is of interest to test the role of in this PIPィイD23ィエD2 System.
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