Urokinase-type Plasminogen Activator (uPA) Promotes Angiogenesis by Attenuating Proline-rich Homeodomain Protein (PRH) Transcription Factor Activity and De-repressing Vascular Endothelial Growth Factor (VEGF) Receptor Expression

Stepanova, Victoria; Jayaraman, Padma-Sheela; Zaitsev, Sergei V.; Lebedeva, Tatiana; Bdeir, Khalil; Kershaw, Rachael; Holman, Kelci R.; Parfyonova, Yelena V.; Semina, Ekaterina V.; Beloglazova, Irina B.; Tkachuk, Vsevolod A.; Cines, Douglas B.

Abstract

Urokinase-type plasminogen activator (uPA) regulates angiogenesis and vascular permeability through proteolytic degradation of extracellular matrix and intracellular signaling initiated upon its binding to uPAR/CD87 and other cell surface receptors. Here, we describe an additional mechanism by which uPA regulates angiogenesis. Ex vivo VEGF-induced vascular sprouting from Matrigel-embedded aortic rings isolated from uPA knock-out (uPA-/-) mice was impaired compared with vessels emanating from wild-type mice. Endothelial cells isolated from uPA-/- mice show less proliferation and migration in response to VEGF than their wild type counterparts or uPA-/- endothelial cells in which expression of wild type uPA had been restored. We reported previously that uPA is transported from cell surface receptors to nuclei through a mechanism that requires its kringle domain. Intranuclear uPA modulates gene transcription by binding to a subset of transcription factors. Here we report that wild type single-chain uPA, but not uPA variants incapable of nuclear transport, increases the expression of cell surface VEGF receptor 1 (VEGFR1) and VEGF receptor 2 (VEGFR2) by translocating to the nuclei of ECs. Intranuclear single-chain uPA binds directly to and interferes with the function of the transcription factor hematopoietically expressed homeodomain protein or proline-rich homeodomain protein (HHEX/ PRH), which thereby lose their physiologic capacity to repress the activity of vehgr1 and vegfr2 gene promoters. These studies identify uPA-dependent de-repression of vegfr1 and vegfr2 gene transcription through binding to HHEX/PRH as a novel mechanism by which uPA mediates the pro-angiogenic effects of VEGF and identifies a potential new target for control of pathologic angiogenesis.

Citation

Stepanova, V., Jayaraman, P.-S., Zaitsev, S. V., Lebedeva, T., Bdeir, K., Kershaw, R., Holman, K. R., Parfyonova, Y. V., Semina, E. V., Beloglazova, I. B., Tkachuk, V. A., & Cines, D. B. (2016). Urokinase-type Plasminogen Activator (uPA) Promotes Angiogenesis by Attenuating Proline-rich Homeodomain Protein (PRH) Transcription Factor Activity and De-repressing Vascular Endothelial Growth Factor (VEGF) Receptor Expression. Journal of Biological Chemistry, 291(29), 15029-15045. https://doi.org/10.1074/jbc.m115.678490

• Journal Article Type: Article
• Acceptance Date: Jul 13, 2015
• Online Publication Date: Jul 15, 2016
• Publication Date: Jul 15, 2016
• Deposit Date: Jan 19, 2021
• Publicly Available Date: Feb 3, 2021
• Journal: Journal of Biological Chemistry
• Print ISSN: 0021-9258
• Electronic ISSN: 1083-351X
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 291
• Issue: 29
• Pages: 15029-15045
• DOI: https://doi.org/10.1074/jbc.m115.678490
• Keywords: Cell Biology; Biochemistry; Molecular Biology
• Public URL: https://nottingham-repository.worktribe.com/output/5246967
• Publisher URL: https://www.jbc.org/article/S0021-9258(20)41251-7/fulltext
• Related Public URLs: https://www.sciencedirect.com/science/article/pii/S0021925820412517
• Additional Information: This article is maintained by: Elsevier; Article Title: Urokinase-type Plasminogen Activator (uPA) Promotes Angiogenesis by Attenuating Proline-rich Homeodomain Protein (PRH) Transcription Factor Activity and De-repressing Vascular Endothelial Growth Factor (VEGF) Receptor Expression; Journal Title: Journal of Biological Chemistry; CrossRef DOI link to publisher maintained version: https://doi.org/10.1074/jbc.M115.678490; Content Type: article; Copyright: © 2016 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.