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Blood. 2015 Mar 19;125(12):1957-67. doi: 10.1182/blood-2014-08-593343. Epub 2014 Dec 23.

Sp1/Sp3 transcription factors regulate hallmarks of megakaryocyte maturation and platelet formation and function.

Author information

1
Department of Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands;
2
Department of Cell Biology, Erasmus MC, Rotterdam, The Netherlands;
3
Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands;
4
Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands;
5
Center for Biomics, and.
6
Proteomics Center, Erasmus MC, Rotterdam, The Netherlands; and.
7
Institute of Molecular Biology and Tumor Research, Philipps-University Marburg, Marburg, Germany.

Abstract

Sp1 and Sp3 belong to the specificity proteins (Sp)/Krüppel-like transcription factor family. They are closely related, ubiquitously expressed, and recognize G-rich DNA motifs. They are thought to regulate generic processes such as cell-cycle and growth control, metabolic pathways, and apoptosis. Ablation of Sp1 or Sp3 in mice is lethal, and combined haploinsufficiency results in hematopoietic defects during the fetal stages. Here, we show that in adult mice, conditional pan-hematopoietic (Mx1-Cre) ablation of either Sp1 or Sp3 has minimal impact on hematopoiesis, whereas the simultaneous loss of Sp1 and Sp3 results in severe macrothrombocytopenia. This occurs in a cell-autonomous manner as shown by megakaryocyte-specific (Pf4-Cre) double-knockout mice. We employed flow cytometry, cell culture, and electron microscopy and show that although megakaryocyte numbers are normal in bone marrow and spleen, they display a less compact demarcation membrane system and a striking inability to form proplatelets. Through megakaryocyte transcriptomics and platelet proteomics, we identified several cytoskeleton-related proteins and downstream effector kinases, including Mylk, that were downregulated upon Sp1/Sp3 depletion, providing an explanation for the observed defects in megakaryopoiesis. Supporting this notion, selective Mylk inhibition by ML7 affected proplatelet formation and stabilization and resulted in defective ITAM receptor-mediated platelet aggregation.

PMID:
25538045
DOI:
10.1182/blood-2014-08-593343
[Indexed for MEDLINE]

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