title: Nesprin-3 regulates endothelial cell morphology, perinuclear cytoskeletal architecture, and flow-induced polarization.
abstract: Changes in blood flow regulate gene expression and protein synthesis in vascular endothelial cells, and this regulation is involved in the development of atherosclerosis. How mechanical stimuli are transmitted from the endothelial luminal surface to the nucleus is incompletely understood. The linker of nucleus and cytoskeleton (LINC) complexes have been proposed as part of a continuous physical link between the plasma membrane and subnuclear structures. LINC proteins nesprin-1, -2, and -4 have been shown to mediate nuclear positioning via microtubule motors and actin. Although nesprin-3 connects intermediate filaments to the nucleus, no functional consequences of nesprin-3 mutations on cellular processes have been described. Here we show that nesprin-3 is robustly expressed in human aortic endothelial cells (HAECs) and localizes to the nuclear envelope. Nesprin-3 regulates HAEC morpho-logy, with nesprin-3 knockdown inducing prominent cellular elongation. Nesprin-3 also organizes perinuclear cytoskeletal organization and is required to attach the centrosome to the nuclear envelope. Finally, nesprin-3 is required for flow-induced polarization of the centrosome and flow-induced migration in HAECs. These results represent the most complete description to date of nesprin-3 function and suggest that nesprin-3 regulates vascular endothelial cell shape, perinuclear cytoskeletal architecture, and important aspects of flow-mediated mechanotransduction.
authors.affiliation: Department of Mechanical and Aerospace Engineering, University of California, Davis, Davis, CA 95616, USA.
authors.forename: Joshua T
authors.initials: JT
authors.lastname: Morgan
authors.forename: Emily R
authors.initials: ER
authors.lastname: Pfeiffer
authors.forename: Twanda L
authors.initials: TL
authors.lastname: Thirkill
authors.forename: Priyadarsini
authors.initials: P
authors.lastname: Kumar
authors.forename: Gordon
authors.initials: G
authors.lastname: Peng
authors.forename: Heidi N
authors.initials: HN
authors.lastname: Fridolfsson
authors.forename: Gordon C
authors.initials: GC
authors.lastname: Douglas
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Starr
authors.forename: Abdul I
authors.initials: AI
authors.lastname: Barakat
mesh_terms: D001011:Aorta / Q000378:metabolism; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D016764:Cell Polarity; D048430:Cell Shape; D002478:Cells, Cultured; D018385:Centrosome / Q000378:metabolism / Q000648:ultrastructure; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism / Q000648:ultrastructure; D040542:Mechanotransduction, Cellular; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009685:Nuclear Envelope / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering
chemical_list: D008565:Membrane Proteins; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; C529565:SYNE3 protein, human