Research Project
Grant-in-Aid for Scientific Research (B)
The goal of this study was to clarify the regulatory mechanisms of thrombopoiesis through analyzing the function of a heterodimeric transcription factor NF-E2.1. Structure and function analysis of small MafA deletion mutant of MafG that lacks 40 amino acids of its C-terminus did not rescue the defective proplatelet formation of megakaryocytes from mafG-null mice when the mutant MafG was supplied from the transgene. This results clearly showed that the MafG C-terminal region is critical to the small Ma function in megakaryocytes. We investigated the proteins interacting to the MafG C-terminal region through the yeast two hybrid screening. We found that NF-E2 p45 interacts with the full length of MafG but not with MafG without the C-terminus. Interestingly, Nrf2, another member of CNC family transcription factor and a related factor of NF-E2 p45, does interact with MafG irrespective of the presence the C-terminal region. These results imply that MafG C-terminal region may contribute to t … More he selection of a heterodimeric partner molecule of MafG.2. Subcellular localization of NF-E2 p45We established transgenic mouse lines expressing Flag-tagged NF-E2 p45 or GFP-fused NF-E2 p45 in megakaryocytes. We also obtained a stable transformant megakaryocytic cell lines expressing Flag-HA-His tagged NF-E2 p45. By using these mice and cells, we are examining the subcellular distribution of NF-E2 p45.3. Target genes of NF-E2 p45We identified a novel gene clone 325 from the subtraction screening of small Maf mutant megakaryocytes and the control megakaryocytes. We disrupted the 325 gene in mice and found that the mice is healthy and fertile displaying the normal platelet count. We are planning to administer anti-platelet antibody and/or myelosupressive reagents to the mice and to see their effects on the thrombopoiesis. We also performed transcriptome analysis of NF-E2 p45-null mice and found that the factors involved in the platelet function and structures are decreased while antioxidant response genes and detoxifying enzyme genes were increased, which suggests the involvement of oxidative stress in the process of megakaryopoiesis and thrombopoiesis. Less
All 2007 2006 2005
All Journal Article (20 results)
Biochem. J. (in press)
Cancer Sci. 98
Pages: 135-139
Biochem.J. (in press)
Mol. Cell. Biol. 26
Pages: 7953-7965
Pages: 4652-4663
Genes Cells 11
Pages: 575-591
Biochem. Biophys. Res. Commun. 339
Pages: 79-88
Mol.Cell.Biol. 26
Biochem.Biophys.Res.Commun. 339
Mol.Cell.Biol. 25
Pages: 8044-8051
J.Biol.Chem. 280
Pages: 4483-4490
J.Immunol. 174
Pages: 2770-2777
Pages: 9360-9368
