title: SMAD1 and SMAD5 Expression Is Coordinately Regulated by FLI1 and GATA2 during Endothelial Development.
abstract: The bone morphogenetic protein (BMP)/SMAD signaling pathway is a critical regulator of angiogenic sprouting and is involved in vascular development in the embryo. SMAD1 and SMAD5, the core mediators of BMP signaling, are vital for this activity, yet little is known about their transcriptional regulation in endothelial cells. Here, we have integrated multispecies sequence conservation, tissue-specific chromatin, in vitro reporter assay, and in vivo transgenic data to identify and validate Smad1+63 and the Smad5 promoter as tissue-specific cis-regulatory elements that are active in the developing endothelium. The activity of these elements in the endothelium was dependent on highly conserved ETS, GATA, and E-box motifs, and chromatin immunoprecipitation showed high levels of enrichment of FLI1, GATA2, and SCL at these sites in endothelial cell lines and E11 dorsal aortas in vivo. Knockdown of FLI1 and GATA2 but not SCL reduced the expression of SMAD1 and SMAD5 in endothelial cells in vitro. In contrast, CD31(+) cKit(-) endothelial cells harvested from embryonic day 9 (E9) aorta-gonad-mesonephros (AGM) regions of GATA2 null embryos showed reduced Smad1 but not Smad5 transcript levels. This is suggestive of a degree of in vivo selection where, in the case of reduced SMAD1 levels, endothelial cells with more robust SMAD5 expression have a selective advantage.
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia The Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.
authors: Jonathon
authors: J
authors: Marks-Bluth
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Anchit
authors: A
authors: Khanna
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Vashe
authors: V
authors: Chandrakanthan
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Julie
authors: J
authors: Thoms
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Thomas
authors: T
authors: Bee
authors: Erasmus MC Stem Cell Institute, Department of Cell Biology, Erasmus University Medical Center, Rotterdam, Netherlands.
authors: Christina
authors: C
authors: Eich
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Young Chan
authors: YC
authors: Kang
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Kathy
authors: K
authors: Knezevic
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia.
authors: Qiao
authors: Q
authors: Qiao
authors: Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom.
authors: Simon
authors: S
authors: Fitch
authors: Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, Maine, USA.
authors: Leif
authors: L
authors: Oxburgh
authors: Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom.
authors: Katrin
authors: K
authors: Ottersbach
authors: Erasmus MC Stem Cell Institute, Department of Cell Biology, Erasmus University Medical Center, Rotterdam, Netherlands The Queen's Medical Research Institute, College of Medicine and Veterinary Medicine, Edinburgh, United Kingdom.
authors: Elaine
authors: E
authors: Dzierzak
authors: The Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom marella.debruijn@imm.ox.ac.uk jpimanda@unsw.edu.au.
authors: Marella F T R
authors: MF
authors: de Bruijn
authors: Lowy Cancer Research Centre and the Prince of Wales Clinical School, The University of New South Wales, Sydney, NSW, Australia Department of Haematology, Prince of Wales Hospital, Sydney, NSW, Australia marella.debruijn@imm.ox.ac.uk jpimanda@unsw.edu.au.
authors: John E
authors: JE
authors: Pimanda
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004727:Endothelium / Q000196:embryology* / Q000378:metabolism; D050989:GATA2 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics / Q000378:metabolism*; D051898:Smad1 Protein / Q000235:genetics*; D051902:Smad5 Protein / Q000235:genetics*
chemical_list: C497426:Fli1 protein, mouse; D050989:GATA2 Transcription Factor; C494712:Gata2 protein, mouse; D051789:Proto-Oncogene Protein c-fli-1; D051898:Smad1 Protein; D051902:Smad5 Protein