# Molecular functions of Tcf-1 in DP thymocytes > **NIH NIH R01** · UNIVERSITY OF CHICAGO · 2022 · $191,994 ## Abstract PROJECT SUMMARY Thymic development is highly responsible for shaping a healthy and balanced T cell immunity. Like other developmental processes it involves coordinated changes in the linear and three-dimensional chromatin organization that allow stage specific transcription events. A central regulator at nearly every stage of T cell differentiation is the DNA binding protein Tcf-1. Studies proposed here will elucidate how Tcf-1 coordinates the action of epigenetic and transcription regulators to instruct the differentiation of CD4+CD8+ DP thymocytes. Tcf- 1 modulates the chromatin landscapes and transcription profiles directly through binding to its conserved DNA sequence, or indirectly in association with other regulatory proteins. DP thymocytes express two forms of Tcf- 1, the full length Tcf-1p45 protein that binds β-catenin, and a short Tcf-1p38 isoform that does not. Lef-1, another member of the Tcf/Lef family of regulators, also expressed in thymocytes has overlapping functions with Tcf-1. In addition Tcf-1 cooperates with the HLH domain DNA binding protein HEB at the DP thymocyte stages through the sharing of ~7000 DNA binding sites genome wide. The presence of both Tcf-1 and HEB at the shared sites is necessary to promote chromatin accessibility and regulate gene transcription. The direct Tcf-1- HEB binding to their conserved motifs in enhancer regions of T cell differentiation genes promotes their expression. By contrast Tcf-1-HEB recruitment to sites lacking conserved motifs, in promoter regions of cell- cycle genes reduces their expression and cell proliferation. Such opposing transcription outcomes likely involve Tcf-1-HEB recruitment to DNA in the context of distinct regulatory complexes. The composition of complexes containing Tcf-1 and HEB, the specific Tcf-1 isoform involved in each complex, and which functions are redundant between Tcf-1 and Lef-1, still remain to be elucidated. The intricate functional co-operation between these factors likely also involves the ability of Tcf-1 and Lef-1 to bend the DNA helix at their binding site which may modulate the 3D chromatin conformation, and define the proximity between co-operating factors and regulatory elements. These findings and the existing literature provide strong premise for the hypothesis that DP thymocyte development is enabled by the cooperation of Tcf-1 isoforms with protein complexes that shape the 3D chromatin structure to differentially regulate gene expression. Two specific aims are proposed. Aim 1 will determine the how Tcf-1p45 and, Tcf-1p33, co-operate with Lef-1 and HEB in the context of distinct regulatory complexes to establish the epigenetic and transcription profile of DP thymocytes and control their developmental progression. Aim 2: will elucidate the roles of Tcf-1p45, Tcf-1p33, Lef-1 with HEB shaping the chromatin conformation, and how this function promotes DP thymocyte development. The proposed studies are expected to highlight a completely new layer in the... ## Key facts - **NIH application ID:** 10287489 - **Project number:** 5R01AI147652-03 - **Recipient organization:** UNIVERSITY OF CHICAGO - **Principal Investigator:** Fotini Gounari - **Activity code:** R01 (R01, R21, SBIR, etc.) - **Funding institute:** NIH - **Fiscal year:** 2022 - **Award amount:** $191,994 - **Award type:** 5 - **Project period:** 2019-11-22 → 2022-02-28 ## Primary source NIH RePORTER: https://reporter.nih.gov/project-details/10287489 ## Citation > US National Institutes of Health, RePORTER application 10287489, Molecular functions of Tcf-1 in DP thymocytes (5R01AI147652-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10287489. Licensed CC0. --- *[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*