The Y14-p53 regulatory circuit in megakaryocyte differentiation and thrombocytopenia.
Thrombocytopenia-absent radius (TAR) syndrome is caused by RBM8A insufficiency. We generated megakaryocyte-specific Rbm8a knockout (Rbm8aKOMK) mice that exhibited marked thrombocytopenia, internal hemorrhage, and splenomegaly, providing evidence that genetic deficiency of Rbm8a causes a disorder of platelet production. Rbm8aKOMK mice accumulated low-ploidy immature megakaryocytes in the bone marrow and exhibited defective platelet activation and aggregation. Accordingly, depletion of Y14 (RBM8A) in human erythroleukemia (HEL) cells compromised phorbol-ester-induced polyploidization. Notably, Y14/RBM8A deficiency induced both p53 and p21 in megakaryocytes and HEL cells. Treatment with a p53 inhibitor restored ex vivo differentiation of Rbm8aKOMK megakaryocytes and unexpectedly activated Y14 expression in HEL cells. Trp53 knockout partially restored megakaryocyte differentiation by reversing cell-cycle arrest and increased platelet counts of Rbm8aKOMK, indicating that excess p53 in part accounts for thrombocytopenia in TAR syndrome. This study provides evidence for the role of the Y14-p53 circuit in platelet production and a potential therapeutic strategy.
Pubmed ID: 34816104
Research resources used in this publication
None foundAdditional research tools detected in this publication
- Novus Biologicals (RRID:SCR_004286)
- PRISM (RRID:SCR_005375)
- Promega (RRID:SCR_006724)
- Thermo Fisher Scientific (RRID:SCR_008452)
- Millipore (RRID:SCR_008983)
- Trimmomatic (RRID:SCR_011848)
- featureCounts (RRID:SCR_012919)
- Abcam (RRID:SCR_012931)
- BD Biosciences (RRID:SCR_013311)
- Bethyl (RRID:SCR_013554)
- MedCalc (RRID:SCR_015044)
- DESeq2 (RRID:SCR_015687)
- clusterProfiler (RRID:SCR_016884)
- HEK293 (RRID:CVCL_0045)
- B6N.FVB-Tg(Pf4-cre/ERT2)3.21Ics/Ics (RRID:MGI:7381137)
Antibodies used in this publication
- Donkey Anti-Rabbit IgG, Whole Ab ECL Antibody, HRP Conjugated (RRID:AB_772206)
- Sheep Anti-Mouse IgG - Horseradish Peroxidase (RRID:AB_772210)
- Mouse Anti-CD41a Monoclonal Antibody, FITC Conjugated, Clone HIP8 (RRID:AB_395858)
- Rat IgG1, kappa Isotype Control, FITC Conjugated, Clone R3-34 (RRID:AB_395508)
- Rat Anti-CD41 Monoclonal Antibody, FITC Conjugated, Clone MWReg30 (RRID:AB_395085)
- Wug.E9 (RRID:AB_2890922)
- PE-labeled Rat Anti-Mouse Integrin αIIbβ3 (GPIIbIIIa, CD41/61) (RRID:AB_2833084)
- GPIX (CD42a) (RRID:AB_2827529)
- GPVI (RRID:AB_2827531)
- gamma H2AX Antibody (RRID:AB_10002815)
- RBM8a Antibody (RRID:AB_2300943)
- Cleaved Caspase-3 (Asp175) Antibody (RRID:AB_2341188)
- CDK1-Specific antibody (RRID:AB_10638617)
- p21 (C-19) (RRID:AB_632126)
- P21 antibody (RRID:AB_2077682)
- P53 antibody (RRID:AB_2206609)
- VWF, VWFpp antibody (RRID:AB_10642840)
- CCNB2 Polyclonal Antibody (RRID:AB_2765068)
- Histone H3 (D1H2) XP Rabbit mAb (RRID:AB_10544537)
- Rabbit Anti-Histone H3, phospho (Ser10) Monoclonal Antibody, Unconjugated, Clone D2C8 (RRID:AB_1549592)
- GAPDH antibody (RRID:AB_2107436)
- Beta Actin antibody (RRID:AB_2289225)
- Y14 antibody [4C4] (RRID:AB_2269494)
Associated grants
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This is a list of tools and resources that we have found mentioned in this publication.
Novus Biologicals (tool)
RRID:SCR_004286
Commercial antibody vendor which supplies antibodies and other products to life science researchers.
View all literature mentionsPRISM (tool)
RRID:SCR_005375
THIS RESOURCE IS NO LONGER IN SERVICE. Documented on May 5,2022.Tool that predicts interactions between transcription factors and their regulated genes from binding motifs. Understanding vertebrate development requires unraveling the cis-regulatory architecture of gene regulation. PRISM provides accurate genome-wide computational predictions of transcription factor binding sites for the human and mouse genomes, and integrates the predictions with GREAT to provide functional biological context. Together, accurate computational binding site prediction and GREAT produce for each transcription factor: 1. putative binding sites, 2. putative target genes, 3. putative biological roles of the transcription factor, and 4. putative cis-regulatory elements through which the factor regulates each target in each functional role.
View all literature mentionsThermo Fisher Scientific (tool)
RRID:SCR_008452
Commercial vendor and service provider of laboratory reagents and antibodies. Supplier of scientific instrumentation, reagents and consumables, and software services.
View all literature mentionsTrimmomatic (tool)
RRID:SCR_011848
Software Java pipeline for trimming tasks for Illumina paired end and single ended data. Flexible Trimmer for Illumina Sequence Data. Pair aware preprocessing tool optimized for Illumina next generation sequencing data. Includes several processing steps for read trimming and filtering. Operating systems Unix/Linux, Mac OS, Windows.
View all literature mentionsfeatureCounts (tool)
RRID:SCR_012919
A read summarization program, which counts mapped reads for the genomic features such as genes and exons.
View all literature mentionsAbcam (tool)
RRID:SCR_012931
A commercial antibody supplier which supplies primary and secondary antibodies, biochemicals, proteins, peptides, lysates, immunoassays and other kits.
View all literature mentionsMedCalc (tool)
RRID:SCR_015044
Statistical software for ROC curve analysis. MedCalc performs several statistical tests such as method comparison, method evaluation, reference intervals, and meta-analysis.
View all literature mentionsDESeq2 (tool)
RRID:SCR_015687
Software package for differential gene expression analysis based on the negative binomial distribution. Used for analyzing RNA-seq data for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates.
View all literature mentionsclusterProfiler (tool)
RRID:SCR_016884
Software R package for statistical analysis and visualization of functional profiles for genes and gene clusters.
View all literature mentionsHEK293 (tool)
RRID:CVCL_0045
Cell line HEK293 is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentionsB6N.FVB-Tg(Pf4-cre/ERT2)3.21Ics/Ics (tool)
RRID:MGI:7381137
laboratory mouse with name B6N.FVB-Tg(Pf4-cre/ERT2)3.21Ics/Ics from MGI.
View all literature mentionsDonkey Anti-Rabbit IgG, Whole Ab ECL Antibody, HRP Conjugated (antibody)
RRID:AB_772206
This unknown targets IgG
View all literature mentionsSheep Anti-Mouse IgG - Horseradish Peroxidase (antibody)
RRID:AB_772210
This monoclonal secondary targets IgG
View all literature mentionsMouse Anti-CD41a Monoclonal Antibody, FITC Conjugated, Clone HIP8 (antibody)
RRID:AB_395858
This monoclonal targets CD41a
View all literature mentionsRat IgG1, kappa Isotype Control, FITC Conjugated, Clone R3-34 (antibody)
RRID:AB_395508
This unknown targets Rat IgG1 kappa Isotype Control FITC Clone R3-34
View all literature mentionsRat Anti-CD41 Monoclonal Antibody, FITC Conjugated, Clone MWReg30 (antibody)
RRID:AB_395085
This monoclonal targets CD41
View all literature mentionsWug.E9 (antibody)
RRID:AB_2890922
This monoclonal targets CD62P (P-selectin)
View all literature mentionsPE-labeled Rat Anti-Mouse Integrin αIIbβ3 (GPIIbIIIa, CD41/61) (antibody)
RRID:AB_2833084
This monoclonal targets integrin αIIbβ31 (GPIIbIIIa, CD41/CD61)
View all literature mentionsGPIX (CD42a) (antibody)
RRID:AB_2827529
This monoclonal targets GPIX (CD42a)
View all literature mentionsgamma H2AX Antibody (antibody)
RRID:AB_10002815
This unknown targets gamma H2AX
View all literature mentionsCleaved Caspase-3 (Asp175) Antibody (antibody)
RRID:AB_2341188
This polyclonal targets Cleaved Caspase-3 (Asp175)
View all literature mentionsCDK1-Specific antibody (antibody)
RRID:AB_10638617
This polyclonal targets CDK1-Specific
View all literature mentionsp21 (C-19) (antibody)
RRID:AB_632126
This unknown targets Raised against a peptide mapping at the carboxy terminus of human p21
View all literature mentionsVWF, VWFpp antibody (antibody)
RRID:AB_10642840
This polyclonal targets VWF, VWFpp
View all literature mentionsCCNB2 Polyclonal Antibody (antibody)
RRID:AB_2765068
This polyclonal targets CCNB2
View all literature mentionsHistone H3 (D1H2) XP Rabbit mAb (antibody)
RRID:AB_10544537
This monoclonal targets Histone H3 (D1H2) XP Rabbit mAb
View all literature mentionsRabbit Anti-Histone H3, phospho (Ser10) Monoclonal Antibody, Unconjugated, Clone D2C8 (antibody)
RRID:AB_1549592
This monoclonal targets Histone H3, phospho (Ser10)
View all literature mentionsGAPDH antibody (antibody)
RRID:AB_2107436
This monoclonal targets GAPDH
View all literature mentionsBeta Actin antibody (antibody)
RRID:AB_2289225
This monoclonal targets Beta Actin
View all literature mentionsY14 antibody [4C4] (antibody)
RRID:AB_2269494
This monoclonal targets RBM8A
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