Expression, activation, and subcellular localization of the Rap1 GTPase in cord blood-derived human megakaryocytes

Experimental Cell Research
Alessandra BalduiniMauro Torti

Abstract

The expression of the small GTPase Rap1 in human megakaryocytes (MKs) differentiated from cord blood (CB)-derived progenitors was investigated. High levels of Rap1 were detected in the majority of mature megakaryocytes independently of days of culture, while a very low percentage of immature megakaryocytes was found to express a small amount of the protein. Rap1 was predominantly detected on internal alpha-granule but not on the plasma membrane. By contrast, CD41 was clearly present on the peripheral plasma membrane, although it also displayed an intracellular localization similar to that of Rap1. Upon thrombin stimulation, both Rap1 and CD41 translocated to the periphery of the cell. At the opposite, RhoA GTPase and glycoprotein Ibalpha were predominantly located at the plasma membrane and did not undergo relocation upon thrombin stimulation. Thrombin induced a dose- and time-dependent activation of Rap1 in mature megakaryocytes. By using a confocal microscopy approach with a specific probe, active Rap1 was detected exclusively at the peripheral plasma membrane. These results demonstrate that expression of Rap1 occurs during maturation rather than differentiation of megakaryocytes from cord blood progenitor cells. Moreover, we...Continue Reading

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Citations

Jun 27, 2014·PLoS Genetics·Patricia Molina-OrtizStéphane Schurmans
Jun 20, 2007·Experimental Hematology·Ian S Hitchcock, Kenneth Kaushansky
Apr 26, 2006·Journal of Thrombosis and Haemostasis : JTH·A BalduiniA Savoia
Oct 12, 2013·Arteriosclerosis, Thrombosis, and Vascular Biology·Jose J LopezRégis Bobe
Sep 16, 2005·Human Molecular Genetics·Alessandro PecciMauro Torti
May 31, 2008·European Journal of Haematology·Che Kang LimLi Sun
Mar 27, 2013·The Journal of Biological Chemistry·Vittorio AbbonanteAlessandra Balduini
May 22, 2018·MicroRNA·Neetu Dahiya, Chintamani D Atreya
Dec 21, 2018·Cell Communication and Signaling : CCS·Sara Ortiz-RiveroCarmen Guerrero
Jan 22, 2005·The Journal of Biological Chemistry·Erika S WittchenKeith Burridge

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