GATA2 at the mitosis-to-G1 transition is critical for definitive hematopoiesis
(2021) 2021 Virtual Annual Scientific Meeting of the ISEH - International Society for Experimental Hematology: ISEH 50th Annual Meeting In Experimental Hematology 100(Suppl). p.35-35- Abstract
- In mitosis, transcription factors (TFs) and RNA polymerase disperse across the cytoplasm leading to transcriptional silencing, but some TFs are retained on condensed chromatin and mark genomic sites, a mechanism termed mitotic bookmarking. In pluripotent and differentiated cells this mechanism is important for pluripotency maintenance, cell reprogramming and lineage inheritance. However, the role of bookmarking in adult stem cells or in an in vivo system is yet to be addressed. Hematopoietic stem cells undergo drastic changes in cell cycle during development while balancing self-renewal and differentiation, suggesting a possible role for bookmarking.
Here, we first addressed the mitotic retention capacity of the hemogenic TFs... (More) - In mitosis, transcription factors (TFs) and RNA polymerase disperse across the cytoplasm leading to transcriptional silencing, but some TFs are retained on condensed chromatin and mark genomic sites, a mechanism termed mitotic bookmarking. In pluripotent and differentiated cells this mechanism is important for pluripotency maintenance, cell reprogramming and lineage inheritance. However, the role of bookmarking in adult stem cells or in an in vivo system is yet to be addressed. Hematopoietic stem cells undergo drastic changes in cell cycle during development while balancing self-renewal and differentiation, suggesting a possible role for bookmarking.
Here, we first addressed the mitotic retention capacity of the hemogenic TFs GATA2, GFI1B and FOS. We show that GATA2 remains bound to chromatin at all phases of cell cycle, as opposed to GFI1B and FOS. The C-terminal zinc finger (C-ZF) and the nuclear localization signal domains are required for GATA2 mitotic binding. Point mutations in the C-ZF associated with leukemia also impact GATA2 retention. To address the role of GATA2-mediated mitotic bookmarking, we have fused GATA2 to a mitosis degradation (MD) domain, which promotes protein destruction at the mitosis-to-G1 transition (M-G1). Degradation of GATA2 at M-G1 impacts the reprogramming of human fibroblasts to hemogenic cells. To address the role of GATA2 at M-G1 in vivo, we have generated a mouse model with the MD domain inserted upstream the Gata2 gene. Remarkably, homozygous mice are lethal, phenocopying Gata2 null mice which die at the onset of definitive hematopoiesis, showing a deficit in hematopoietic stem and progenitor cells.
These findings implicate GATA2 as a mitotic bookmarking factor and its critical role at M-G1 for definitive hematopoiesis. Overall, our study highlights a dependency on mitotic bookmarkers for in vivo lineage commitment. (Less)
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https://lup.lub.lu.se/record/ed1132b5-d1c4-49db-bf1b-ba5fa1aba5ed
- author
- Alves, Rita LU ; Haider, Jakob LU ; Thelaus, Louise LU ; Lindgren, Aida ; Ferreira, Gabriela LU ; Rosa, Fábio LU ; Gonzalez, Javier and Pereira, Carlos-Filipe LU
- organization
-
- LUCC: Lund University Cancer Centre
- Division of Molecular Medicine and Gene Therapy
- Cell Reprogramming in Hematopoiesis and Immunity (research group)
- StemTherapy: National Initiative on Stem Cells for Regenerative Therapy
- WCMM-Wallenberg Centre for Molecular Medicine
- Heparin bindning protein in cardiothoracic surgery (research group)
- Translational Sepsis research (research group)
- Stem Cell Center
- publishing date
- 2021-08-01
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Experimental Hematology
- volume
- 100
- issue
- Suppl
- pages
- 35 - 35
- publisher
- Elsevier
- conference name
- 2021 Virtual Annual Scientific Meeting of the ISEH - International Society for Experimental Hematology: ISEH 50th Annual Meeting
- conference dates
- 2021-08-25 - 2021-08-28
- ISSN
- 1873-2399
- DOI
- 10.1016/j.exphem.2021.12.381
- language
- English
- LU publication?
- yes
- id
- ed1132b5-d1c4-49db-bf1b-ba5fa1aba5ed
- date added to LUP
- 2023-08-10 21:26:53
- date last changed
- 2023-08-14 14:50:09
@misc{ed1132b5-d1c4-49db-bf1b-ba5fa1aba5ed, abstract = {{In mitosis, transcription factors (TFs) and RNA polymerase disperse across the cytoplasm leading to transcriptional silencing, but some TFs are retained on condensed chromatin and mark genomic sites, a mechanism termed mitotic bookmarking. In pluripotent and differentiated cells this mechanism is important for pluripotency maintenance, cell reprogramming and lineage inheritance. However, the role of bookmarking in adult stem cells or in an in vivo system is yet to be addressed. Hematopoietic stem cells undergo drastic changes in cell cycle during development while balancing self-renewal and differentiation, suggesting a possible role for bookmarking.<br/><br/>Here, we first addressed the mitotic retention capacity of the hemogenic TFs GATA2, GFI1B and FOS. We show that GATA2 remains bound to chromatin at all phases of cell cycle, as opposed to GFI1B and FOS. The C-terminal zinc finger (C-ZF) and the nuclear localization signal domains are required for GATA2 mitotic binding. Point mutations in the C-ZF associated with leukemia also impact GATA2 retention. To address the role of GATA2-mediated mitotic bookmarking, we have fused GATA2 to a mitosis degradation (MD) domain, which promotes protein destruction at the mitosis-to-G1 transition (M-G1). Degradation of GATA2 at M-G1 impacts the reprogramming of human fibroblasts to hemogenic cells. To address the role of GATA2 at M-G1 in vivo, we have generated a mouse model with the MD domain inserted upstream the Gata2 gene. Remarkably, homozygous mice are lethal, phenocopying Gata2 null mice which die at the onset of definitive hematopoiesis, showing a deficit in hematopoietic stem and progenitor cells.<br/><br/>These findings implicate GATA2 as a mitotic bookmarking factor and its critical role at M-G1 for definitive hematopoiesis. Overall, our study highlights a dependency on mitotic bookmarkers for in vivo lineage commitment.}}, author = {{Alves, Rita and Haider, Jakob and Thelaus, Louise and Lindgren, Aida and Ferreira, Gabriela and Rosa, Fábio and Gonzalez, Javier and Pereira, Carlos-Filipe}}, issn = {{1873-2399}}, language = {{eng}}, month = {{08}}, note = {{Conference Abstract}}, number = {{Suppl}}, pages = {{35--35}}, publisher = {{Elsevier}}, series = {{Experimental Hematology}}, title = {{GATA2 at the mitosis-to-G1 transition is critical for definitive hematopoiesis}}, url = {{http://dx.doi.org/10.1016/j.exphem.2021.12.381}}, doi = {{10.1016/j.exphem.2021.12.381}}, volume = {{100}}, year = {{2021}}, }