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:The t(12;21) chromosomal translocation, targeting the gene encoding the RUNX1 transcription factor, is observed in 25% of pediatric acute lymphoblastic leukemia (ALL) and is an initiating event in the disease. To elucidate the mechanism by which RUNX1 disruption initiates leukemogenesis, we investigated its normal role in murine B-cell development. Gene expression analysis and genome-wide Runx1-occupancy studies support the hypothesis that Runx1 reinforces the transcription factor network in B-cell progenitors governing early B-cell survival and development . 1) To identify Runx1 target genes in early B-cell progenitors, we used a whole genome microarray for analyzing the gene expression profile in a proB-cell line [BMiFLT3(15-3)] engineered to overexpresses the Runx1 transcription factor fused to the ligand binding domain of the estrogen receptor. The use of an inducible Runx1 fusion protein allowed specific translocation of Runx1 into the nucleus activation of Runx1 target genes. 2) To identify Runx1 target genes in early B-cell progenitors, we used a whole genome microarray for analyzing the gene expression profiles of proB-cells (CD19+/B220med/CD93+) isolated from Runx1+/+Cd79ahCre/+ and Runx1fl/flCd79ahCre/+ mice. Two independent experiments were peformed.