title: Polarity Protein Scrib Facilitates Endothelial Inflammatory Signaling.
abstract: OBJECTIVE
  The polarity protein Scrib is highly expressed in endothelial cells and is required for planar cell polarity. Scrib also facilitates recycling of integrin α5 to the plasma membrane. Because integrin α5 signals the presence of the inflammatory matrix protein fibronectin, we hypothesized that Scrib contributes to endothelial inflammatory signaling.
  
  
  RESULTS
  Cytokine treatment of human umbilical vein endothelial cells induced an inflammatory response as evident by the induction of vascular cell adhesion molecule-1 (VCAM-1). Downregulation of Scrib greatly attenuated this effect. In endothelial-specific conditional Scrib knockout mice, in vivo lipopolysaccharide treatment resulted in an impaired VCAM-1 induction. These effects were functionally relevant because Scrib small interfering RNAs in human umbilical vein endothelial cells attenuated the VCAM-1-mediated leukocyte adhesion in response to tumor necrosis factor-α. In vivo, tamoxifen-induced endothelial-specific deletion of Scrib resulted in a reduced VCAM-1-mediated leukocyte adhesion in response to tumor necrosis factor-α in the mouse cremaster model. This effect was specific for Scrib and not mediated by other polarity proteins. Moreover, it did not involve integrin α5 or classic pathways supporting inflammatory signaling, such as nuclear factor κ light chain enhancer of activated B-cells or MAP kinases. Co-immunoprecipitation/mass spectrometry identified the zinc finger transcription factor GATA-like protein-1 as a novel Scrib interacting protein. Small interfering RNA depletion of GATA-like protein-1 decreased the tumor necrosis factor-α-stimulated VCAM-1 induction to a similar extent as loss of Scrib did. Silencing of Scrib reduced GATA-like protein-1 protein, but not mRNA abundance.
  
  
  CONCLUSIONS
  Scrib is a novel proinflammatory regulator in endothelial cells, which maintains the protein expression of GATA-like protein-1.
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Christoph
authors.initials: C
authors.lastname: Kruse
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Angela R M
authors.initials: AR
authors.lastname: Kurz
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Katalin
authors.initials: K
authors.lastname: Pálfi
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Patrick O
authors.initials: PO
authors.lastname: Humbert
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Markus
authors.initials: M
authors.lastname: Sperandio
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.). brandes@vrc.uni-frankfurt.de.
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: Christian
authors.initials: C
authors.lastname: Fork
authors.affiliation: From the Institute for Cardiovascular Physiology, Goethe University, Frankfurt, Germany (C.K., K.P., R.P.B., C.F., U.R.M.); Walter-Brendel Center of Experimental Medicine, Ludwig-Maximilians University, Munich, Germany (A.R.M.K., M.S.); Cell Cycle and Cancer Genetics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, and Sir Peter MacCallum Department of Oncology, Department of Pathology, Department of Molecular Biology and Biochemistry, The University of Melbourne, Parkville, Victoria, Australia (P.O.H.); and DZHK (German Centre for Cardiovascular Research), partner sites Rhine-Main and Munich, Germany (C.K., A.R.M.K., K.P., M.S., R.P.B., C.F., U.R.M.).
authors.forename: U Ruth
authors.initials: UR
authors.lastname: Michaelis
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002339:Carotid Arteries / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D050982:GATA1 Transcription Factor / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D047908:Intracellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D050982:GATA1 Transcription Factor; D047908:Intracellular Signaling Peptides and Proteins; D012313:RNA; C494689:Gata1 protein, mouse; C508433:scrib protein, mouse
keywords: GATA-like protein-1; VCAM-1; cell polarity; endothelial cells; inflammation