title: CXXC5 is a transcriptional activator of Flk-1 and mediates bone morphogenic protein-induced endothelial cell differentiation and vessel formation.
abstract: CXXC5 is a member of a small subset of proteins containing CXXC-type zinc-finger domain. Here, we show that CXXC5 is a transcription factor activating Flk-1, a receptor for vascular endothelial growth factor. CXXC5 and Flk-1 were accumulated in nucli and membrane of mouse embryonic stem cells (mESCs), respectively, during their endothelial differentiation. CXXC5 overexpression induced Flk-1 transcription in both endothelium-differentiated mESCs and human umbilical vein endothelial cells (HUVECs). In vitro DNA binding assay showed direct interaction of CXXC5 on the Flk-1 promoter region, and mutation on its DNA-binding motif abolished transcriptional activity. We showed that bone morphorgenetic protein 4 (BMP4) induced CXXC5 transcription in the cells, and inhibitors of BMP signaling suppressed the CXXC5 induction and the consequent Flk-1 induction by BMP4 treatment. CXXC5 knockdown resulted in suppression of BMP4-induced stress fiber formation (56.8 ± 1.3% decrease, P<0.05) and migration (54.6 ± 1.9% decrease, P<0.05) in HUVECs. The in vivo roles of CXXC5 in BMP-signaling-specific vascular development and angiogenesis were shown by specific defect of caudal vein plex vessel formation (57.9 ± 11.8% decrease, P<0.05) in cxxc5 morpholino-injected zebrafish embryos and by suppression of BMP4-induced angiogenesis in subcutaneously injected Matrigel plugs in CXXC5(-/-) mice. Overall, CXXC5 is a transcriptional activator for Flk-1, mediating BMP signaling for differentiation and migration of endothelial cell and vessel formation.
authors.affiliation: 2Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, 120-749, South Korea. kychoi@yonsei.ac.kr.
authors.forename: Hyun-Yi
authors.initials: HY
authors.lastname: Kim
authors.forename: Dong-Hwa
authors.initials: DH
authors.lastname: Yang
authors.forename: Song-Weon
authors.initials: SW
authors.lastname: Shin
authors.forename: Mi-Yeon
authors.initials: MY
authors.lastname: Kim
authors.forename: Jae-Hyun
authors.initials: JH
authors.lastname: Yoon
authors.forename: Suhyun
authors.initials: S
authors.lastname: Kim
authors.forename: Hae-Chul
authors.initials: HC
authors.lastname: Park
authors.forename: Dong Woo
authors.initials: DW
authors.lastname: Kang
authors.forename: Dosik
authors.initials: D
authors.lastname: Min
authors.forename: Man-Wook
authors.initials: MW
authors.lastname: Hur
authors.forename: Kang-Yell
authors.initials: KY
authors.lastname: Choi
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins; D006801:Humans; D051379:Mice; D014157:Transcription Factors; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D043925:Angiogenesis Inducing Agents; D055415:Bone Morphogenetic Protein 4; C540448:CXXC5 protein, human; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: HUVECs; caudal vein plex vessel formation; mouse embryonic stem cells