MicroRNAs 10a and 10b Regulate the Expression of Human Platelet Glycoprotein Ibα for Normal Megakaryopoiesis
Abstract
:1. Introduction
2. Results
2.1. Web-Based Prediction of miRNAs Targeting the Human GP Ibα mRNA
2.2. miRNA 10a and 10b Regulate Human GP Ibα Expression
3. Discussion
4. Materials and Methods
4.1. Prediction of Putative miRNA Binding Sites in the 3′-UTR of Human GP1BA Gene
4.2. Generation of GP1BA 3′-UTR Reporter Constructs and Site-Directed Mutagenesis
4.3. Cell Lines
4.4. Luciferase Assays
4.5. Flow Cytometry Analysis for Transient Expression of GP Ibα in GP Ibβ/IX-Expressing Chinese Hamster Ovary (CHO) Cells
4.6. In Vitro Culture and Transfection of Murine Lineage Negative Cells
4.7. Reverse Transcription Polymerase Chain Reaction (RT-PCR)
4.8. Quantitative Polymerase Chain Reaction (qPCR)
4.9. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
3′-UTR | Three prime untranslated region |
BSS | Bernard-Soulier Syndrome |
GP | Glycoprotein |
vWf | Von Willebrand factor |
FBS | Fetal bovine serum |
RT-PCR | Reverse Transcription Polymerase Chain Reaction |
qPCR | Quantitative Polymerase Chain Reaction |
CHO | Chinese Hamster Ovary |
IMDM | Iscove′s Modified Dulbecco′s Medium |
FITC | Fluorescein isothiocyanate |
RBC | Red blood cell |
GATA | GATA binding protein |
Ets | E26 transformation-specific sequence family transcriptional factor |
Meis1 | Meis homeobox 1 |
MAFB | V-maf musculoaponeurotic fibrosarcoma oncogene homolog B |
c-Myb | Avian myeloblastosis virus oncogene cellular homolog |
NF-E2 | Nuclear factor, erythroid-derived 2 |
HoxB4 | Homeobox B4 |
HoxD4 | Homeobox D4 |
References
- López, J.A.; Andrews, R.K.; Afshar-Kharghan, V.; Berndt, M.C. Bernard-Soulier Syndrome. Blood 1998, 91, 4397–4418. [Google Scholar] [PubMed]
- Kanaji, T.; Russell, S.; Cunningham, J.; Izuhara, K.; Fox, J.E.; Ware, J. Megakaryocyte proliferation and ploidy regulated by the cytoplasmic tail of glycoprotein Ibα. Blood 2004, 104, 3161–3168. [Google Scholar] [CrossRef] [PubMed]
- Lepage, A.; Leboeuf, M.; Cazenave, J.P.; de la, S.C.; Lanza, F.; Uzan, G. The α(IIb)β(3) integrin and GPIb-V-IX complex identify distinct stages in the maturation of CD34+ cord blood cells to megakaryocytes. Blood 2000, 96, 4169–4177. [Google Scholar] [PubMed]
- Poujol, C.; Ware, J.; Nieswandt, B.; Nurden, A.T.; Nurden, P. Absence of GPIbα is responsible for aberrant membrane development during megakaryocyte maturation: Ultrastructural study using a transgenic model. Exp. Hematol. 2002, 30, 352–360. [Google Scholar] [CrossRef]
- Takahashi, R.; Sekine, N.; Nakatake, T. Influence of monoclonal antiplatelet glycoprotein antibodies on in vitro human megakaryocyte colony formation and proplatelet formation. Blood 1999, 93, 1951–1958. [Google Scholar] [PubMed]
- Nurden, P.; Chretien, F.; Poujol, C.; Winckler, J.; Borel-Derlon, A.; Nurden, A. Platelet ultrastructural abnormalities in three patients with type 2B von Willebrand disease. Br. J. Haematol. 2000, 110, 704–714. [Google Scholar] [CrossRef] [PubMed]
- Jurak Begonja, A.; Hoffmeister, K.M.; Hartwig, J.H.; Falet, H. FlnA-null megakaryocytes prematurely release large and fragile platelets that circulate poorly. Blood 2011, 118, 2285–2295. [Google Scholar] [CrossRef] [PubMed]
- Hashimoto, Y.; Ware, J. Identification of essential GATA and Ets binding motifs within the promoter of the platelet glycoprotein Ibα gene. J. Biol. Chem. 1995, 270, 24532–24539. [Google Scholar] [CrossRef] [PubMed]
- Prandini, M.H.; Martin, F.; Thevenon, D.; Uzan, G. The tissue-specific transcriptional regulation of the megakaryocytic glycoprotein IIb gene is controlled by interactions between a repressor and positive cis-acting elements. Blood 1996, 88, 2062–2070. [Google Scholar] [PubMed]
- Bastian, L.S.; Yagi, M.; Chan, C.; Roth, G.J. Analysis of the megakaryocyte glycoprotein IX promoter identifies positive and negative regulatory domains and functional GATA and Ets sites. J. Biol. Chem. 1996, 271, 18554–18560. [Google Scholar] [CrossRef] [PubMed]
- Lepage, A.; Uzan, G.; Touche, N.; Morales, M.; Cazenave, J.P.; Lanza, F.; de la, S.C. Functional characterization of the human platelet glycoprotein V gene promoter: A specific marker of late megakaryocytic differentiation. Blood 1999, 94, 3366–3380. [Google Scholar] [PubMed]
- Ambros, V. The functions of animal microRNAs. Nature 2004, 431, 350–355. [Google Scholar] [CrossRef] [PubMed]
- Barroga, C.F.; Pham, H.; Kaushansky, K. Thrombopoietin regulates c-Myb expression by modulating micro RNA 150 expression. Exp. Hematol. 2008, 36, 1585–1592. [Google Scholar] [CrossRef] [PubMed]
- Lu, J.; Guo, S.; Ebert, B.L.; Zhang, H.; Peng, X.; Bosco, J.; Pretz, J.; Schlanger, R.; Wang, J.Y.; Mak, R.H.; et al. MicroRNA-mediated control of cell fate in megakaryocyte-erythrocyte progenitors. Dev. Cell 2008, 14, 843–853. [Google Scholar] [CrossRef] [PubMed]
- Garzon, R.; Pichiorri, F.; Palumbo, T.; Iuliano, R.; Cimmino, A.; Aqeilan, R.; Volinia, S.; Bhatt, D.; Alder, H.; Marcucci, G.; et al. MicroRNA fingerprints during human megakaryocytopoiesis. Proc. Natl. Acad. Sci. USA 2006, 103, 5078–5083. [Google Scholar] [CrossRef] [PubMed]
- Edelstein, L.C.; Bray, P.F. MicroRNAs in platelet production and activation. Blood 2011, 117, 5289–5296. [Google Scholar] [CrossRef] [PubMed]
- Edelstein, L.C.; McKenzie, S.E.; Shaw, C.; Holinstat, M.A.; Kunapuli, S.P.; Bray, P.F. MicroRNAs in platelet production and activation. J. Thromb. Haemost. 2013, 11, 340–350. [Google Scholar] [CrossRef] [PubMed]
- Romania, P.; Lulli, V.; Pelosi, E.; Biffoni, M.; Peschle, C.; Marziali, G. MicroRNA 155 modulates megakaryopoiesis at progenitor and precursor level by targeting Ets-1 and Meis1 transcription factors. Br. J. Haematol. 2008, 143, 570–580. [Google Scholar] [CrossRef] [PubMed]
- Dangwal, S.; Thum, T. MicroRNAs in platelet biogenesis and function. Thromb. Haemost. 2012, 108, 599–604. [Google Scholar] [CrossRef] [PubMed]
- Ware, J.; Russell, S.R.; Marchese, P.; Ruggeri, Z.M. Expression of human platelet Glycoprotein Ibα in transgenic mice. J. Biol. Chem. 1993, 268, 8376–8382. [Google Scholar] [PubMed]
- Zhou, H.; Ran, Y.; Da, Q.; Shaw, T.S.; Shang, D.; Reddy, A.K.; Lopez, J.A.; Ballantyne, C.M.; Ware, J.; Wu, H.; et al. Defective association of the platelet Glycoprotein Ib-IX complex with the Glycosphingolipid-enriched membrane domain inhibits murine thrombus and atheroma formation. J. Immunol. 2016, 197, 288–295. [Google Scholar] [CrossRef] [PubMed]
- Hall, M.A.; Curtis, D.J.; Metcalf, D.; Elefanty, A.G.; Sourris, K.; Robb, L.; Gothert, J.R.; Jane, S.M.; Begley, C.G. The critical regulator of embryonic hematopoiesis, SCL, is vital in the adult for megakaryopoiesis, erythropoiesis, and lineage choice in CFU-S12. Proc. Natl. Acad. Sci. USA 2003, 100, 992–997. [Google Scholar] [CrossRef] [PubMed]
- Hart, A.; Melet, F.; Grossfeld, P.; Chien, K.; Jones, C.; Tunnacliffe, A.; Favier, R.; Bernstein, A. Fli-1 is required for murine vascular and megakaryocytic development and is hemizygously deleted in patients with thrombocytopenia. Immunity 2000, 13, 167–177. [Google Scholar] [CrossRef]
- Metcalf, D.; Carpinelli, M.R.; Hyland, C.; Mifsud, S.; Dirago, L.; Nicola, N.A.; Hilton, D.J.; Alexander, W.S. Anomalous megakaryocytopoiesis in mice with mutations in the c-Myb gene. Blood 2005, 105, 3480–3487. [Google Scholar] [CrossRef] [PubMed]
- Zhong, Y.; Sullenbarger, B.; Lasky, L.C. Effect of increased HoxB4 on human megakaryocytic development. Biochem. Biophys. Res. Commun. 2010, 398, 377–382. [Google Scholar] [CrossRef] [PubMed]
- Lecine, P.; Blank, V.; Shivdasani, R. Characterization of the hematopoietic transcription factor NF-E2 in primary murine megakaryocytes. J. Biol. Chem. 1998, 273, 7572–7578. [Google Scholar] [CrossRef] [PubMed]
- Wu, L.; Belasco, J.G. Let me count the ways: Mechanisms of gene regulation by miRNAs and siRNAs. Mol. Cell 2008, 29, 1–7. [Google Scholar] [CrossRef] [PubMed]
- Valinezhad, O.A.; Safaralizadeh, R.; Kazemzadeh-Bavili, M. Mechanisms of miRNA-mediated gene regulation from common downregulation to mRNA-specific upregulation. Int. J. Genom. 2014, 2014, 970607. [Google Scholar] [CrossRef] [PubMed]
- TargetScanHuman—Prediction of MicroRNA Targets. Available online: http://www.targetscan.org (accessed on 13 August 2012).
- MicroRNA.org—Targets and Expression. Available online: http://www.microrna.org/microrna (accessed on 13 August 2012).
- Segal Lab of Computational Biology. Available online: http://genie.weizmann.ac.il/pubs/mir07/mir07_dyn_data.html (accessed on 13 August 2012).
- López, J.A.; Leung, B.; Reynolds, C.C.; Li, C.Q.; Fox, J.E.B. Efficient plasma membrane expression of a functional platelet glycoprotein Ib-IX complex requires the presence of its three subunits. J. Biol. Chem. 1992, 267, 12851–12859. [Google Scholar] [PubMed]
- Peng, Y.; Berndt, M.C.; Cruz, M.A.; Lopez, J.A. The α1 Helix-β13 strand spanning Leu214-Val229 of platelet glycoprotein Ibα facilitates the interaction with von Willebrand factor: Evidence from characterization of the epitope of monoclonal antibody AP1. Blood 2004, 104, 3971–3978. [Google Scholar] [CrossRef] [PubMed]
- Nishikii, H.; Eto, K.; Tamura, N.; Hattori, K.; Heissig, B.; Kanaji, T.; Sawaguchi, A.; Goto, S.; Ware, J.; Nakauchi, H. Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells. J. Exp. Med. 2008, 205, 1917–1927. [Google Scholar] [CrossRef] [PubMed]
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Zhang, Z.; Ran, Y.; Shaw, T.S.; Peng, Y. MicroRNAs 10a and 10b Regulate the Expression of Human Platelet Glycoprotein Ibα for Normal Megakaryopoiesis. Int. J. Mol. Sci. 2016, 17, 1873. https://doi.org/10.3390/ijms17111873
Zhang Z, Ran Y, Shaw TS, Peng Y. MicroRNAs 10a and 10b Regulate the Expression of Human Platelet Glycoprotein Ibα for Normal Megakaryopoiesis. International Journal of Molecular Sciences. 2016; 17(11):1873. https://doi.org/10.3390/ijms17111873
Chicago/Turabian StyleZhang, Zuping, Yali Ran, Tanner S. Shaw, and Yuandong Peng. 2016. "MicroRNAs 10a and 10b Regulate the Expression of Human Platelet Glycoprotein Ibα for Normal Megakaryopoiesis" International Journal of Molecular Sciences 17, no. 11: 1873. https://doi.org/10.3390/ijms17111873