title: Cooperative Stimulation of Megakaryocytic Differentiation by Gfi1b Gene Targets Kindlin3 and Talin1.
abstract: Understanding the production and differentiation of megakaryocytes from progenitors is crucial for realizing the biology and functions of these vital cells. Previous gene ablation studies demonstrated the essential role of the transcriptional repressor Gfi1b (growth factor independence 1b) in the generation of both erythroid and megakaryocytic cells. However, our recent work has demonstrated the down-regulation of this factor during megakaryocytic differentiation. In this study we identify two new gene targets of Gfi1b, the cytoskeletal proteins Kindlin3 and Talin1, and demonstrate the inverse expression and functions of these cytoskeletal targets relative to Gfi1b, during megakaryocytic differentiation. Both kindlin3 and talin1 promoters exhibit dose dependent Gfi1b and LSD1 (lysine specific demethylase 1; a Gfi1b cofactor) enrichment in megakaryocytes and repression in non-hematopoietic cells. Accordingly the expression of these genes is elevated in gfi1b mutant and LSD1 inhibited hematopoietic cells, while during megakaryocytic differentiation, declining Gfi1b levels fostered the reciprocal upregulation of these cytoskeletal factors. Concordantly, manipulation of Kindlin3 and Talin1 expression demonstrated positive correlation with megakaryocytic differentiation with over-expression stimulating, and inhibition diminishing, this process. Co-operativity between these factors and integrins in promoting differentiation was further underscored by physical interactions between them and integrinβ3/CD61 and by stimulation of differentiation by the Talin1 head domain, which is necessary and sufficient for integrin activation. Therefore this study demonstrates the significance of Gfi1b regulated Kindlin3-Talin1 expression in driving megakaryocytic differentiation and highlights the contribution of cytoskeletal agents in the developmental progression of these platelet progenitors.
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Divya
authors: D
authors: Singh
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Ghanshyam
authors: G
authors: Upadhyay
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Ananya
authors: A
authors: Sengupta
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Mohammed A
authors: MA
authors: Biplob
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Shaleen
authors: S
authors: Chakyayil
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Tiji
authors: T
authors: George
authors: Department of Biology, The City College of New York and The Graduate Center of The City University of New York, 160 Convent Avenue, New York, NY, 10031, United States of America.
authors: Shireen
authors: S
authors: Saleque
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D006801:Humans; D039661:Integrin beta3 / Q000378:metabolism; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D016608:Talin / Q000378:metabolism*
chemical_list: C477322:FERMT3 protein, human; C111977:GFI1B protein, human; D039661:Integrin beta3; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; C518832:TLN1 protein, human; D016608:Talin