title: Sox factors transcriptionally regulate ROBO4 gene expression in developing vasculature in zebrafish.
abstract: Despite their importance as members of the Roundabout (Robo) family in the control of axonal and vascular patterning, the transcriptional regulation of these genes is poorly understood. In this study, we show that members of the Sry-related high mobility box (Sox) transcription factor family as being transcriptional regulators of roundabout4 (robo4), a Robo gene family member that participates in sprouting angiogenesis in vivo, in zebrafish. Double whole mount in situ hybridization analysis in zebrafish embryos revealed co-localization of the vascular relevant Sox factors sox7 or sox18 mRNA with robo4 transcripts in developing intersomitic vessels. A 3-kb human ROBO4 promoter element was able to drive reporter expression in zebrafish to recapitulate the endogenous temporal intersomitic vessel expression pattern of robo4. EMSA analysis confirmed binding of Sox18 to a canonical Sox binding site (from -1170 bp to -1176 bp) in the ROBO4 promoter (3 kb), and mutation analysis indicated that this site was partially responsible for ROBO4 promoter activity in ECs. A combination of gain- and loss-of-function analysis identified Sox7 and Sox18 co-regulation of robo4 but not fli1a transcripts in zebrafish. Finally, Sox-mediated robo4 transcriptional regulation is conserved across evolution. These studies imply Sox-mediated transcriptional regulation of Robo4 in the developing embryonic vasculature.
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Ganesh V
authors: GV
authors: Samant
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Marcus O
authors: MO
authors: Schupp
authors: Institute for Molecular Bioscience, the University of Queensland, Brisbane, Queensland 4072, Australia.
authors: Mathias
authors: M
authors: François
authors: Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Milan 26-20133, Italy.
authors: Silvia
authors: S
authors: Moleri
authors: Department of Medicine and Kidney Disease Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Rajendra K
authors: RK
authors: Kothinti
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Chang Zoon
authors: CZ
authors: Chun
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Indranil
authors: I
authors: Sinha
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Suzanna
authors: S
authors: Sellars
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Noah
authors: N
authors: Leigh
authors: Prescient Life Sciences, New Delhi 122 002, India.
authors: Kallal
authors: K
authors: Pramanik
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Mark A
authors: MA
authors: Horswill
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Indulekha
authors: I
authors: Remadevi
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Keguo
authors: K
authors: Li
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: George A
authors: GA
authors: Wilkinson
authors: Department of Medicine and Kidney Disease Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226.
authors: Niloofar M
authors: NM
authors: Tabatabai
authors: Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Milan 26-20133, Italy.
authors: Monica
authors: M
authors: Beltrame
authors: Institute for Molecular Bioscience, the University of Queensland, Brisbane, Queensland 4072, Australia.
authors: Peter
authors: P
authors: Koopman
authors: Division of Developmental Biology, Developmental Vascular Biology Program, Department of Pediatrics, Children's Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin 53226. Electronic address: rramchan@mcw.edu.
authors: Ramani
authors: R
authors: Ramchandran
mesh_terms: D000818:Animals; D002465:Cell Movement; D004252:DNA Mutational Analysis; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D051379:Mice; D009154:Mutation; D009389:Neovascularization, Pathologic; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000502:physiology; D055760:SOXF Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000096:biosynthesis* / Q000502:physiology; D000097905:Roundabout Proteins
chemical_list: D011956:Receptors, Cell Surface; D055760:SOXF Transcription Factors; D029961:Zebrafish Proteins; C502398:ROBO4 protein, zebrafish; D000097905:Roundabout Proteins; C095473:SOX18 protein, human; C484456:SOX7 protein, human