|
| Status |
Public on Dec 18, 2014 |
| Title |
Sp1/Sp3 transcription factors regulate hallmarks of megakaryocyte maturation, and platelet formation and function |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
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 ablation of either Sp1 or Sp3 has minimal
impact on hematopoiesis, while the simultaneous loss of Sp1 and Sp3 results in severe
macrothrombocytopenia and platelet dysfunction. 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. We show that Mylk is required for proplatelet formation and
stabilization and for ITAM-receptor mediated platelet aggregation. Our data highlights
the specific vs generic role of these ubiquitous transcription factors in the highly
specialized megakaryocytic lineage.
|
| |
|
| Overall design |
Megakaryocyte mRNA profiles of Sp1fl/fl::Sp3fl/fl (WTlox) and Pf4-Cre::Sp1fl/fl::Sp3fl/fl (dKO) mice were generated by deep sequencing, in triplicate.
|
| |
|
| Contributor(s) |
Kulu DI, van IJcken WF, van de Werken HJ, Philipsen S |
| Citation(s) |
25538045 |
| |
| Submission date |
Jun 20, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Harmen Jan George van de Werken |
| E-mail(s) |
h.vandewerken@erasmusmc.nl
|
| Phone |
+31-10-7038906
|
| Organization name |
Erasmus MC
|
| Department |
Urology
|
| Lab |
Cancer Computational Biology Center
|
| Street address |
Wytemaweg 80
|
| City |
Rotterdam |
| ZIP/Postal code |
3015 CN |
| Country |
Netherlands |
| |
|
| Platforms (1) |
| GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
|
| Samples (6)
|
|
| Relations |
| BioProject |
PRJNA253207 |
| SRA |
SRP043469 |
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