Study of molecular mechanisms of CD4 silencing and role s of Runx proteins during lymphocyte development
Grant-in-Aid for Scientific Research (B)
|Allocation Type||Single-year Grants|
|Research Institution||RIKEN (Institute of Physical and Chemical Research)(2004)|
TANIUCHI Ichiro RIKEN (Institute of Physical and Chemical Research), Laboratory for transcriptional regulation, Team leader, 免疫転写制御研究チーム, チームリーダー (20284573)
畠山 鎮次 九州大学, 生体防御医学研究所, 助教授 (70294973)
嘉村 巧 九州大学, 生体防御医学研究所, 助教授 (40333455)
中山 敬一 九州大学, 生体防御医学研究所, 教授 (80291508)
中山 啓子 東北大学, 大学院・医学研究科, 教授 (60294972)
|Project Period (FY)
2003 – 2004
Completed(Fiscal Year 2004)
|Budget Amount *help
¥15,300,000 (Direct Cost : ¥15,300,000)
Fiscal Year 2004 : ¥7,400,000 (Direct Cost : ¥7,400,000)
Fiscal Year 2003 : ¥7,900,000 (Direct Cost : ¥7,900,000)
|Keywords||epigenetics / CD4 gene / gene silencing / Runx family / エピジェネティクス / Runxファミリィー / CD4 / T細胞分化 / Runx / silencing|
The aim of this research is to understand the molecular mechanisms of CD4 silencing and function of Runx transcriptional factors during lymphocyte development. Because a main experimental approach was in vivo analyses of gene manipulated mice, we have generated several mutant mouse strains including gene targeted mice and transgenic mice. By using gene targeting in Es cells and Cre-loxP system, we have generated flox mutant allele for Runx1,Runx3 or CBFβ gene for conditional inactivation by induction of Cre recombinase by tissue and stage specific manner. We have also generated Runx3 transgenic mouse in which myc-epitope tagged Runx3 was specifically expressed in all αβT lineage cells. Another mutant strain, the Runx1^<Δ446> mouse, in which VWRPY amino acid sequence of Runx1 protein was deleted, was kindly provided. By analyzing these mutant mice, we have obtained following results.
1.iNKT cells are originated from DP thymocytes and require Runx1 function for their development.
nd Runx3 possess redundant function in CD4 silencing and thymocyte maturation. In the double knock out of Runx1 and Runx3, positive selection and thymocyte maturation were severely impaired.
3.The C-terminal VWRPY sequence of Runx1 is essential for CD4 silencing in both immature DN thymocytes and mature CD8-lineage thymocytes. However, VWRPY independent transcriptional repression operate ob another target genes of Runx1. Moreover, the VWRPY motif is essential for Runx1 function in iNKT cells development.
4.Expression of Runx3 by transgene is not sufficient to initiate CD4 silencing. Runx3 would not a molecule that defines lineage specificity of CD4 silencing.
5.CBFβ plays essential roles for Runx complex function during thymocytes differentiation.
6.Binding of Runx1 to three Runx recognition motifs in Eβ enhancer at TCRβ locus is essential for activation of TCRβ gene. However, mutation on one Runx motif out of three result in partial impairment of TCRβ activation in DN thymocytes, but did not affect maintenance of TCRβ activation in mature T cells Less
Research Products (18results)