PML::RARA and GATA2 proteins interact via DNA templates to induce aberrant self- renewal in mouse and human hematopoietic cells

• 成果类型: Article
• 署名作者: Katerndahl, Casey D. S.; Rogers, Olivia R. S.; Day, Ryan B.; Xu, Ziheng; Helton, Nichole M.; Ramakrishnan, Sai Mukund; Miller, Christopher A.; Ley, Timothy J.
• 署名单位: Washington University (WUSTL)
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-11625
• DOI: 10.1073/pnas.2317690121
• 发表日期: 2024-04-30
• 关键词: acute promyelocytic leukemia; retinoic acid receptor; pml-rar-alpha; acute myeloid-leukemia; pml/rar-alpha; transcription factor; histone deacetylase; differentiation block; response element; cross-talk

摘要

The underlying mechanism(s) by which the PML::RARA fusion protein initiates acute promyelocytic leukemia is not yet clear. We defined the genomic binding sites of PML::RARA in primary mouse and human hematopoietic progenitor cells with V5-tagged PML::RARA, using anti-V5-PML::RARA chromatin immunoprecipitation sequencing and CUT&RUN approaches. Most genomic PML::RARA binding sites were found in regions that were already chromatin-accessible (defined by ATAC-seq) in unmanipulated, wild-type promyelocytes, suggesting that these regions are open prior to PML::RARA expression. We found that GATA binding motifs, and the direct binding of the chromatin pioneering factor GATA2, were significantly enriched near PML::RARA binding sites. Proximity labeling studies revealed that PML::RARA interacts with similar to 250 proteins in primary mouse hematopoietic cells; GATA2 and 33 others require PML::RARA binding to DNA for the interaction to occur, suggesting that binding to their cognate DNA target motifs may stabilize their interactions. In the absence of PML::RARA , Gata2 overexpression induces many of the same epigenetic and transcriptional changes as PML::RARA . These findings suggested that PML::RARA may indirectly initiate its transcriptional program by activating Gata2 expression: Indeed, we demonstrated that inactivation of Gata2 prior to PML::RARA expression prevented its ability to induce self-renewal. These data suggested that GATA2 binding creates accessible chromatin regions enriched for both GATA and Retinoic Acid Receptor Element motifs, where GATA2 and PML::RARA can potentially bind and interact with each other. In turn, PML::RARA binding to DNA promotes a feed-forward transcriptional program by positively regulating Gata2 expression. Gata2 may therefore be required for PML::RARA to establish its transcriptional program.