통찰 - Computational Biology - # EWSR1::FLI1 fusion protein regulation of chromatin structure and gene expression in Ewing sarcoma

The Critical Role of the DBD-α4 Helix in EWSR1::FLI1-Mediated Genome Regulation and Chromatin Restructuring in Ewing Sarcoma

Q: How might the DBD-α4 helix mechanistically stabilize the cooperative binding of EWSR1::FLI1 at GGAA microsatellites, such as through direct DNA interactions, interactions with other proteins, or intramolecular interactions with the EWSR1 domain?

The DBD-α4 helix likely stabilizes the cooperative binding of EWSR1::FLI1 at GGAA microsatellites through various mechanisms. One possible mechanism is through direct DNA interactions, where the DBD-α4 helix directly interacts with the GGAA repeats in the DNA, facilitating stable binding of the fusion protein to these specific sites. This direct interaction could involve specific amino acid residues in the DBD-α4 helix that recognize and bind to the GGAA motifs, enhancing the affinity and stability of the binding. Another potential mechanism could involve interactions with other proteins. The DBD-α4 helix may interact with other transcription factors or chromatin regulators that are involved in the binding and regulation of GGAA microsatellites. These protein-protein interactions could stabilize the binding of EWSR1::FLI1 at these sites and modulate the transcriptional activity of the fusion protein. Additionally, intramolecular interactions with the EWSR1 domain could play a role in stabilizing the binding of EWSR1::FLI1 at GGAA microsatellites. The DBD-α4 helix may interact with the EWSR1 domain within the fusion protein, leading to conformational changes that enhance the binding affinity to GGAA repeats. This intramolecular interaction could contribute to the overall stability and function of EWSR1::FLI1 in regulating gene expression at these specific genomic loci.

Q: What are the potential therapeutic implications of targeting the DBD-α4 helix or EWSR1::FLI1-mediated transcription hubs for the treatment of Ewing sarcoma?

Targeting the DBD-α4 helix or EWSR1::FLI1-mediated transcription hubs could have significant therapeutic implications for the treatment of Ewing sarcoma. By disrupting the cooperative binding of EWSR1::FLI1 at GGAA microsatellites, potential therapeutic strategies could aim to inhibit the oncogenic activity of the fusion protein and prevent the dysregulation of gene expression that drives Ewing sarcoma tumorigenesis. Specifically targeting the DBD-α4 helix could disrupt the formation of transcription hubs and alter the 3D chromatin organization, leading to the downregulation of oncogenic pathways and the inhibition of tumor growth. Inhibiting the binding of EWSR1::FLI1 at GGAA microsatellites could prevent the formation of de-novo enhancers and the activation of genes involved in proliferation, migration, and invasion pathways, which are critical for the oncogenic transformation of Ewing sarcoma cells. Furthermore, targeting EWSR1::FLI1-mediated transcription hubs could offer a more precise and targeted approach to disrupting the oncogenic activity of the fusion protein. By disrupting the formation of these hubs, it may be possible to selectively inhibit the expression of genes that are essential for Ewing sarcoma tumorigenesis, while sparing normal cellular functions.

Q: How do the findings from this study on the role of the DBD-α4 helix in chromatin restructuring and gene regulation relate to the broader understanding of how oncogenic fusion proteins reprogram the epigenome and 3D genome organization in other cancer types?

The findings from this study provide valuable insights into how oncogenic fusion proteins, such as EWSR1::FLI1, reprogram the epigenome and 3D genome organization in Ewing sarcoma, which can be extrapolated to understand similar mechanisms in other cancer types driven by fusion proteins. The role of the DBD-α4 helix in stabilizing the binding of EWSR1::FLI1 at GGAA microsatellites and promoting the formation of transcription hubs highlights a common mechanism by which oncogenic fusion proteins modulate gene expression and chromatin architecture. This mechanism may be applicable to other fusion proteins in different cancer types, where the fusion protein binds to specific genomic loci and drives oncogenic transcriptional programs. Furthermore, the study's emphasis on the importance of cooperative binding at specific genomic sites and the impact on 3D chromatin organization underscores a general principle of how oncogenic fusion proteins reshape the epigenome and regulate gene expression in cancer. Understanding these mechanisms can provide insights into potential therapeutic strategies that target the dysregulated gene expression and chromatin architecture driven by oncogenic fusion proteins in various cancer types.

핵심 개념

The DBD-α4 helix of the EWSR1::FLI1 fusion protein is crucial for its cooperative binding at GGAA microsatellites, which underlies the formation of topologically associated domains (TADs), chromatin loops, enhancers, and productive transcription hubs that drive oncogenic gene expression in Ewing sarcoma.

초록

This study investigates the mechanism by which the DBD-α4 helix of the EWSR1::FLI1 fusion protein promotes transcription and oncogenic transformation in Ewing sarcoma. The authors used a multi-omics approach, including RNA-seq, CUT&Tag, and Micro-C, to assess chromatin organization, active chromatin marks, genome binding, and gene expression in Ewing sarcoma cells expressing EWSR1::FLI1 constructs with and without the DBD-α4 helix.

The key findings are:

The DBD-α4 helix is required for EWSR1::FLI1 to restructure 3D chromatin organization, including the formation of TADs and chromatin loops.
The DBD-α4 helix facilitates cooperative binding of EWSR1::FLI1 at GGAA microsatellites, particularly those that are longer and denser in GGAA motifs. This binding underlies the formation of TADs, chromatin loops, and enhancers that drive Ewing-specific gene expression.
The DBD-α4 helix is necessary for the formation of productive transcription hubs at key Ewing sarcoma genes, such as FCGRT and CCND1, by promoting EWSR1::FLI1 binding at dense GGAA microsatellites and enabling chromatin looping and enhancer activation.
These findings were largely consistent between the A-673 and TTC-466 Ewing sarcoma cell lines, underscoring the critical role of the DBD-α4 helix in EWSR1::FLI1-mediated genome regulation and oncogenic transformation.

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biorxiv.org

통계

The EWSR1::FLI1 fusion protein is the only genetic alteration in Ewing sarcoma tumors.
Ewing sarcoma has a 5-year survival rate of only 10-30% for metastatic and relapsed patients.
The authors detected 14,326 and 65 differentially interacting regions (DIRs) in DBD+ and DBD cells, respectively, compared to knockdown (KD) cells in A-673 cells.
In TTC-466 cells, the authors detected 1,742 DIRs in DBD+ cells and 72 DIRs in DBD cells compared to KD cells.

인용구

"The DBD-α4 helix is crucial for cooperative binding of EWSR1::FLI1 at GGAA microsatellites. This binding underlies many aspects of genome regulation by EWSR1::FLI1 such as formation of TADs, chromatin loops, enhancers and productive transcription hubs."
"The DBD-α4 helix facilitates formation of short-range loops by promoting binding at long and dense GGAA microsatellites, thus leading to more gene regulation."
"The DBD-α4 helix is required for modulating the enhancer landscape in Ewing cells to specifically promote oncogenic transformation."

핵심 통찰 요약

The DBD-α4 helix of EWSR1::FLI1 is required for GGAA microsatellite binding that underlies genome regulation in Ewing sarcoma

by Bayanjargal,... 게시일 www.biorxiv.org 01-31-2024

https://www.biorxiv.org/content/10.1101/2024.01.31.578127v2

더 깊은 질문

How might the DBD-α4 helix mechanistically stabilize the cooperative binding of EWSR1::FLI1 at GGAA microsatellites, such as through direct DNA interactions, interactions with other proteins, or intramolecular interactions with the EWSR1 domain?

The DBD-α4 helix likely stabilizes the cooperative binding of EWSR1::FLI1 at GGAA microsatellites through various mechanisms. One possible mechanism is through direct DNA interactions, where the DBD-α4 helix directly interacts with the GGAA repeats in the DNA, facilitating stable binding of the fusion protein to these specific sites. This direct interaction could involve specific amino acid residues in the DBD-α4 helix that recognize and bind to the GGAA motifs, enhancing the affinity and stability of the binding.
Another potential mechanism could involve interactions with other proteins. The DBD-α4 helix may interact with other transcription factors or chromatin regulators that are involved in the binding and regulation of GGAA microsatellites. These protein-protein interactions could stabilize the binding of EWSR1::FLI1 at these sites and modulate the transcriptional activity of the fusion protein.
Additionally, intramolecular interactions with the EWSR1 domain could play a role in stabilizing the binding of EWSR1::FLI1 at GGAA microsatellites. The DBD-α4 helix may interact with the EWSR1 domain within the fusion protein, leading to conformational changes that enhance the binding affinity to GGAA repeats. This intramolecular interaction could contribute to the overall stability and function of EWSR1::FLI1 in regulating gene expression at these specific genomic loci.

What are the potential therapeutic implications of targeting the DBD-α4 helix or EWSR1::FLI1-mediated transcription hubs for the treatment of Ewing sarcoma?

Targeting the DBD-α4 helix or EWSR1::FLI1-mediated transcription hubs could have significant therapeutic implications for the treatment of Ewing sarcoma. By disrupting the cooperative binding of EWSR1::FLI1 at GGAA microsatellites, potential therapeutic strategies could aim to inhibit the oncogenic activity of the fusion protein and prevent the dysregulation of gene expression that drives Ewing sarcoma tumorigenesis.
Specifically targeting the DBD-α4 helix could disrupt the formation of transcription hubs and alter the 3D chromatin organization, leading to the downregulation of oncogenic pathways and the inhibition of tumor growth. Inhibiting the binding of EWSR1::FLI1 at GGAA microsatellites could prevent the formation of de-novo enhancers and the activation of genes involved in proliferation, migration, and invasion pathways, which are critical for the oncogenic transformation of Ewing sarcoma cells.
Furthermore, targeting EWSR1::FLI1-mediated transcription hubs could offer a more precise and targeted approach to disrupting the oncogenic activity of the fusion protein. By disrupting the formation of these hubs, it may be possible to selectively inhibit the expression of genes that are essential for Ewing sarcoma tumorigenesis, while sparing normal cellular functions.

How do the findings from this study on the role of the DBD-α4 helix in chromatin restructuring and gene regulation relate to the broader understanding of how oncogenic fusion proteins reprogram the epigenome and 3D genome organization in other cancer types?

The findings from this study provide valuable insights into how oncogenic fusion proteins, such as EWSR1::FLI1, reprogram the epigenome and 3D genome organization in Ewing sarcoma, which can be extrapolated to understand similar mechanisms in other cancer types driven by fusion proteins.
The role of the DBD-α4 helix in stabilizing the binding of EWSR1::FLI1 at GGAA microsatellites and promoting the formation of transcription hubs highlights a common mechanism by which oncogenic fusion proteins modulate gene expression and chromatin architecture. This mechanism may be applicable to other fusion proteins in different cancer types, where the fusion protein binds to specific genomic loci and drives oncogenic transcriptional programs.
Furthermore, the study's emphasis on the importance of cooperative binding at specific genomic sites and the impact on 3D chromatin organization underscores a general principle of how oncogenic fusion proteins reshape the epigenome and regulate gene expression in cancer. Understanding these mechanisms can provide insights into potential therapeutic strategies that target the dysregulated gene expression and chromatin architecture driven by oncogenic fusion proteins in various cancer types.