Project description:Cellular binary fate decisions require the progeny to silence genes associated with the alternative fate. The major subsets of alpha:beta T cells have been extensively studied as a model system for fate decisions. While the transcription factor RUNX3 is required for the initiation of Cd4 silencing in CD8 T cell progenitors, it is not required to maintain the silencing of Cd4 and other helper T lineage genes. The other runt domain containing protein, RUNX1, silences Cd4 in an earlier T cell progenitor, but this silencing is reversed whereas the gene silencing after RUNX3 expression is not reverse. Therefore, we hypothesized that RUNX3 and not RUNX1 recruits other factors that maintains the silencing of helper T lineage genes in CD8 T cells. To this end, we performed a proteomics screen of RUNX1 and RUNX3 to determine candidate silencing factors.
Project description:Series of samples studying effect of knocking out RUNX1 (a.k.a. AML1) in mouse embryos at 8.5 and 12 days. Wildtype and knockout animals at the two time points were generated and all conditions done in duplicate.
Project description:Series of samples studying effect of knocking out RUNX1 (a.k.a. AML1) in mouse embryos at 8.5 and 12 days. Wildtype and knockout animals at the two time points were generated and all conditions done in duplicate. Keywords: other
Project description:To clarify the role of AML1-ETO in aberrant hematopoiesis, we generated conditional AML1-ETO knock-in mice. Our research indicated AML1-ETO induction impaired bone marrow hematopoietic reconstitution and resulted in aberrant accumulation of Lin-Sca-1+c-Kit+ (LSK) cells. To clarify the potential mechanism, we compared gene expression profiling and epigenetic alterations of LSK cells between AML1/ETO mouse (conditional knock-in of Runx1-Runx1t1; Mx1-Cre mouse model) and the control mouse (AML1/ETO mice without Mx1-Cre).
Project description:To clarify the role of AML1-ETO in aberrant hematopoiesis, we generated conditional AML1-ETO knock-in mice. Our research indicated AML1-ETO induction impaired bone marrow hematopoietic reconstitution and resulted in aberrant accumulation of Lin-Sca-1+c-Kit+ (LSK) cells. To clarify the potential mechanism, we compared gene expression profiling and epigenetic alterations of LSK cells between AML1/ETO mouse (conditional knock-in of Runx1-Runx1t1; Mx1-Cre mouse model) and the control mouse (AML1/ETO mice without Mx1-Cre).
Project description:Human histone deacetylase 3 (HDAC3) plays an important role in gene transcription in diseased human cells, such as leukemia. The t(8;21) chromosomal translocation is one of the most commonly observed genetic abnormalities associated with acute myeloid leukemia. This translocation generates the AML1-ETO fusion protein between the wild-type RUNX1 transcription factor and wild-type ETO transcriptional corepressor. To better understand the role of HDAC3 in t(8;21) leukemogenesis, the human HDAC3-containing complexes were isolated from stably-transfected HeLa cells by using anti-FLAG immunoprecipitation. The resulting complexes were resolved in SDS-PAGE. The components of the complexes were identified using LC-MS/MS. We report here that the human RUNX1 transcription is a component of the HDAC3 complexes. We demonstrate that HDAC3 and RUNX1 collaboratively repress AML1-ETO-mediated transcription. These results reveal new insight into how AML1-ETO, RUNX1, and HDAC3 crosstalk to deregulate gene transcription in t(8;21) leukemia cells.
