title: A Unique Role for Endothelial Cell Kinesin Light Chain 1, Variant 1 in Leukocyte Transendothelial Migration.
abstract: A reservoir of parajunctional membrane in endothelial cells, the lateral border recycling compartment (LBRC), is critical for transendothelial migration (TEM). We have previously shown that targeted recycling of the LBRC to the site of TEM requires microtubules and a kinesin molecular motor. However, the identity of the kinesin and mechanism of cargo binding were not known. We show that microinjection of endothelial cells with a monoclonal antibody specific for kinesin-1 significantly blocked LBRC-targeted recycling and TEM. In complementary experiments, knocking down KIF5B, a ubiquitous kinesin-1 isoform, in endothelial cells significantly decreased targeted recycling of the LBRC and leukocyte TEM. Kinesin heavy chains move cargo along microtubules by one of many kinesin light chains (KLCs), which directly bind the cargo. Knocking down KLC 1 isoform variant 1 (KLC1C) significantly decreased LBRC-targeted recycling and TEM, whereas knocking down other isoforms of KLC1 had no effect. Re-expression of KLC1C resistant to the knockdown shRNA restored targeted recycling and TEM. Thus kinesin-1 and KLC1C are specifically required for targeted recycling and TEM. These data suggest that of the many potential combinations of the 45 kinesin family members and multiple associated light chains, KLC1C links the LBRC to kinesin-1 (KIF5B) during targeted recycling and TEM. Thus, KLC1C can potentially be used as a target for anti-inflammatory therapy.
authors.affiliation: Department of Pathology, The Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
authors.forename: Bita F
authors.initials: BF
authors.lastname: Cyrus
authors.affiliation: Department of Pathology, The Feinberg School of Medicine, Northwestern University, Chicago, Illinois. Electronic address: wamuller@northwestern.edu.
authors.forename: William A
authors.initials: WA
authors.lastname: Muller
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D007249:Inflammation / Q000503:physiopathology / Q000517:prevention & control; D016547:Kinesins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D007962:Leukocytes / Q000502:physiology*; D008845:Microinjections; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D008870:Microtubules / Q000502:physiology; D017354:Point Mutation / Q000235:genetics; D014161:Transduction, Genetic; D057705:Transendothelial and Transepithelial Migration / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; C500506:KIF5A protein, human; C488101:KIF5B protein, human; C000715688:KLC1 protein, human; D008869:Microtubule-Associated Proteins; D016547:Kinesins