Control of endothelial tubulogenesis by Rab and Ral GTPases, and apical targeting of caveolin-1-labeled vacuoles
Here, we examine known GTPase regulators of vesicle trafficking events to assess whether they affect endothelial cell (EC) lumen and tube formation. We identify novel roles for the small GTPases Rab3A, Rab3B, Rab8A, Rab11A, Rab27A, RalA, RalB and caveolin-1 in co-regulating membrane trafficking even...
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| description | Here, we examine known GTPase regulators of vesicle trafficking events to assess whether they affect endothelial cell (EC) lumen and tube formation. We identify novel roles for the small GTPases Rab3A, Rab3B, Rab8A, Rab11A, Rab27A, RalA, RalB and caveolin-1 in co-regulating membrane trafficking events that control EC lumen and tube formation. siRNA suppression of individual GTPases such as Rab3A, Rab8A, and RalB markedly inhibit tubulogenesis, while greater blockade is observed with combinations of siRNAs such as Rab3A and Rab3B, Rab8A and Rab11A, and RalA and RalB. These combinations of siRNAs also disrupt very early events in lumen formation including the formation of intracellular vacuoles. In contrast, knockdown of the endocytosis regulator, Rab5A, fails to inhibit EC tube formation. Confocal microscopy and real-time videos reveal that caveolin-1 strongly labels intracellular vacuoles and localizes to the EC apical surface as they fuse to form the luminal membrane. In contrast, Cdc42 and Rab11A localize to a perinuclear, subapical region where intracellular vacuoles accumulate and fuse during lumen formation. Our new data demonstrates that EC tubulogenesis is coordinated by a series of small GTPases to control polarized membrane trafficking events to generate, deliver, and fuse caveolin-1-labeled vacuoles to create the apical membrane surface. |
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Our new data demonstrates that EC tubulogenesis is coordinated by a series of small GTPases to control polarized membrane trafficking events to generate, deliver, and fuse caveolin-1-labeled vacuoles to create the apical membrane surface.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0235116</identifier><identifier>PMID: 32569321</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adenoviruses ; Biology and Life Sciences ; Caveolin ; Caveolin 1 - metabolism ; Caveolin-1 ; Caveolins ; Cdc42 protein ; Cell Membrane - metabolism ; Circulatory system ; Collagen ; Collagen - metabolism ; Confocal microscopy ; Embryonic development ; Endocytosis ; Endothelial cells ; Endothelium ; Enzymes ; Exocytosis ; Green Fluorescent Proteins - metabolism ; GTPases ; Guanosine triphosphatases ; Guanosinetriphosphatase ; Health aspects ; Human Umbilical Vein Endothelial Cells - metabolism ; Humans ; Intracellular ; Membrane trafficking ; Membranes ; Microscopy ; Models, Biological ; Morphogenesis ; Pharmacology ; Physiological aspects ; Physiology ; Protein Transport ; rab GTP-Binding Proteins - metabolism ; Rab3A protein ; rac GTP-Binding Proteins - metabolism ; ral GTP-Binding Proteins - metabolism ; Regulators ; RNA, Small Interfering - metabolism ; siRNA ; src-Family Kinases - metabolism ; Vacuoles ; Vacuoles - metabolism</subject><ispartof>PloS one, 2020-06, Vol.15 (6), p.e0235116-e0235116</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Norden et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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metabolism</topic><topic>Caveolin-1</topic><topic>Caveolins</topic><topic>Cdc42 protein</topic><topic>Cell Membrane - metabolism</topic><topic>Circulatory system</topic><topic>Collagen</topic><topic>Collagen - metabolism</topic><topic>Confocal microscopy</topic><topic>Embryonic development</topic><topic>Endocytosis</topic><topic>Endothelial cells</topic><topic>Endothelium</topic><topic>Enzymes</topic><topic>Exocytosis</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>GTPases</topic><topic>Guanosine triphosphatases</topic><topic>Guanosinetriphosphatase</topic><topic>Health aspects</topic><topic>Human Umbilical Vein Endothelial Cells - metabolism</topic><topic>Humans</topic><topic>Intracellular</topic><topic>Membrane trafficking</topic><topic>Membranes</topic><topic>Microscopy</topic><topic>Models, Biological</topic><topic>Morphogenesis</topic><topic>Pharmacology</topic><topic>Physiological aspects</topic><topic>Physiology</topic><topic>Protein Transport</topic><topic>rab GTP-Binding Proteins - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Norden, Pieter R</au><au>Sun, Zheying</au><au>Davis, George E</au><au>Wary, Kishore K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Control of endothelial tubulogenesis by Rab and Ral GTPases, and apical targeting of caveolin-1-labeled vacuoles</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2020-06-22</date><risdate>2020</risdate><volume>15</volume><issue>6</issue><spage>e0235116</spage><epage>e0235116</epage><pages>e0235116-e0235116</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Here, we examine known GTPase regulators of vesicle trafficking events to assess whether they affect endothelial cell (EC) lumen and tube formation. We identify novel roles for the small GTPases Rab3A, Rab3B, Rab8A, Rab11A, Rab27A, RalA, RalB and caveolin-1 in co-regulating membrane trafficking events that control EC lumen and tube formation. siRNA suppression of individual GTPases such as Rab3A, Rab8A, and RalB markedly inhibit tubulogenesis, while greater blockade is observed with combinations of siRNAs such as Rab3A and Rab3B, Rab8A and Rab11A, and RalA and RalB. These combinations of siRNAs also disrupt very early events in lumen formation including the formation of intracellular vacuoles. In contrast, knockdown of the endocytosis regulator, Rab5A, fails to inhibit EC tube formation. Confocal microscopy and real-time videos reveal that caveolin-1 strongly labels intracellular vacuoles and localizes to the EC apical surface as they fuse to form the luminal membrane. In contrast, Cdc42 and Rab11A localize to a perinuclear, subapical region where intracellular vacuoles accumulate and fuse during lumen formation. Our new data demonstrates that EC tubulogenesis is coordinated by a series of small GTPases to control polarized membrane trafficking events to generate, deliver, and fuse caveolin-1-labeled vacuoles to create the apical membrane surface.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>32569321</pmid><doi>10.1371/journal.pone.0235116</doi><tpages>e0235116</tpages><orcidid>https://orcid.org/0000-0002-1549-1610</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Adenoviruses Biology and Life Sciences Caveolin Caveolin 1 - metabolism Caveolin-1 Caveolins Cdc42 protein Cell Membrane - metabolism Circulatory system Collagen Collagen - metabolism Confocal microscopy Embryonic development Endocytosis Endothelial cells Endothelium Enzymes Exocytosis Green Fluorescent Proteins - metabolism GTPases Guanosine triphosphatases Guanosinetriphosphatase Health aspects Human Umbilical Vein Endothelial Cells - metabolism Humans Intracellular Membrane trafficking Membranes Microscopy Models, Biological Morphogenesis Pharmacology Physiological aspects Physiology Protein Transport rab GTP-Binding Proteins - metabolism Rab3A protein rac GTP-Binding Proteins - metabolism ral GTP-Binding Proteins - metabolism Regulators RNA, Small Interfering - metabolism siRNA src-Family Kinases - metabolism Vacuoles Vacuoles - metabolism |
| title | Control of endothelial tubulogenesis by Rab and Ral GTPases, and apical targeting of caveolin-1-labeled vacuoles |
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