LDB1-COMPLEXES · Role of Ldb1 binding partners Eto2 and cdk9 during stem cells development and erythroid differentiation
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2006-11-01 → 2008-10-31
- EU contribution
- €156,624
- Participants
- 1
- Scheme
- EIF
Lines connect the coordinator with its partners. CORDIS does not always give exact coordinates for projects before 2014. These points are placed at city or country level.
Results in brief
Final Activity Report Summary - Ldb1-complexes (Role of Ldb1 binding partners Eto2 and cdk9 during stem cells development and erythroid differentiation)
The emergence of the hematopoietic system, comprising all blood cells, is tightly controlled by a set of highly conserved transcription factors. The way in which these factors regulate blood cells formation, proliferation and differentiation is largely unknown. The aim of the present work was to analyse the functions of two newly identified key transcription factors, namely Eto2 and Cdk9. A proteomic screen was conducted in the laboratory to identify Eto2 and Cdk9 associated proteins in erythroid cells in order to better understand their functions. This screen showed that Eto2 and Cdk9 could form different sub-complexes, each of them being connected to different chromatin remodelling factors. Relevant data remained unpublished by the time of the project completion. In combination to this proteomic analysis, a genome-wide mapping of Eto2 binding site was carried out using the chromatin immunoprecipitation sequencing (ChIP-sequencing) technology, which allowed for the identification of the deoxyribonucleic acid (DNA) elements and target genes bound by the different Eto2 complexes. The same experiments were also underway with Cdk9. Furthermore, identical experiments were successfully conducted with other hematopoietic transcription factors connected to Eto2 and Cdk9, such as Ldb1, Tal1 and Gata1. The collected results shed light on how these transcription factors co-regulated the proliferating capacity of erythroid progenitors. Combining all these results would finally allow us to unravel how transcription factor networks were formed and to control cell fate during hematopoietic tissue development.
Data: CORDIS, © European Union
Project objective
Ldb1, a ubiquitously expressed LIM domain binding protein, is essential in a number of tissues during development. It interacts with Gata1, Tal1, E2A and Lmo2 to form a transcription factor complex regulating late erythroid genes.It has been identified by the host laboratory a number of novel Ldb1 interacting proteins in erythroleukemic cells, in particular the repressor protein Eto2 (and its family member Mtgr1) and the cyclin dependent kinase Cdk9. Morpholino mediated knockdowns in zebrafish show these f actors to be essential for definitive hematopoiesis. In accordance with the zebrafish results these factors are co-expressed in pre-hematopoietic cells of the early mouse embryo although the complexes were originally identified in late erythroid cells.Based on the change in subcellular localisation of Eto2, we postulate that it plays a central role in the transition from the migration and expansion phase of the pre-heamatopoietic cells to the establishment of definitive hematopoietic stem cells. My project will focus on the understanding of the role of the two newly identified Ldb1 binding partners, (Eto2 and Cdk9) in the regulation of the formation of the blood system.In order to achieve my goals, I will use a multidisciplinary approach involving affinity tagging of the proteins (allowing purification and mass spectrometry identification of interacting partners), functional testing of the novel partner proteins in zebrafish, and mouse genetic modification to obtain in vivo data through conditional knockouts.The role of Eto2 and Cdk9 will be studied at the cellular and organism level. I will use mice and cells to identify their target genes and to gain insight into their role in the development of stem cells and erythroid differentiation in vivo.
Original text from CORDIS.
Participants
- ERASMUS MC · ROTTERDAMCoordinatorCity levelNetherlands
Links
Data: CORDIS, © European Union