title: Syk associates with clathrin and mediates phosphatidylinositol 3-kinase activation during human rhinovirus internalization.
abstract: Human rhinovirus (HRV) causes the common cold. The most common acute infection in humans, HRV is a leading cause of exacerbations of asthma and chronic obstruction pulmonary disease because of its ability to exacerbate airway inflammation by altering epithelial cell biology upon binding to its receptor, ICAM-1. ICAM-1 regulates not only viral entry and replication but also signaling pathways that lead to inflammatory mediator production. We recently demonstrated the Syk tyrosine kinase to be an important mediator of HRV-ICAM-1 signaling: Syk regulates replication-independent p38 MAPK activation and IL-8 expression. In leukocytes, Syk regulates receptor-mediated internalization via PI3K. Although PI3K has been shown to regulate HRV-induced IL-8 expression and clathrin-mediated endocytosis of HRV, the role of airway epithelial Syk in this signaling pathway is not known. We postulated that Syk regulates PI3K activation and HRV endocytosis in the airway epithelium. Using confocal microscopy and immunoprecipitation, we demonstrated recruitment of the normally cytosolic Syk to the plasma membrane upon HRV16-ICAM-1 binding, along with Syk-clathrin coassociation. Subsequent incubation at 37 degrees C to permit internalization revealed redistribution of Syk to punctate structures resembling endosomes and colocalization with HRV16. Internalized HRV was not detected in cells overexpressing the kinase inactive Syk(K396R) mutant, indicating that kinase activity was necessary for endocytosis. HRV-induced PI3K activation was dependent on Syk; Syk knockdown by small interfering RNA significantly decreased phosphorylation of the PI3K substrate Akt. Together, these data reveal Syk to be an important mediator of HRV endocytosis and HRV-induced PI3K activation.
authors.affiliation: Division of Respirology, Multi-Organ Transplantation Programme, University Health Network, Department of Medicine, University of Toronto, Ontario, Canada.
authors.forename: Christine
authors.initials: C
authors.lastname: Lau
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wang
authors.forename: Lihua
authors.initials: L
authors.lastname: Song
authors.forename: Michelle
authors.initials: M
authors.lastname: North
authors.forename: Shahina
authors.initials: S
authors.lastname: Wiehler
authors.forename: David
authors.initials: D
authors.lastname: Proud
authors.forename: Chung-Wai
authors.initials: CW
authors.lastname: Chow
mesh_terms: D001980:Bronchi / Q000378:metabolism / Q000821:virology*; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000821:virology; D002966:Clathrin / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012229:Rhinovirus / Q000502:physiology*; D000072377:Syk Kinase; D053586:Virus Internalization; D000099149:Ezrin
chemical_list: D002966:Clathrin; D003598:Cytoskeletal Proteins; D018799:Intercellular Adhesion Molecule-1; D047908:Intracellular Signaling Peptides and Proteins; D019869:Phosphatidylinositol 3-Kinases; D011505:Protein-Tyrosine Kinases; D051057:Proto-Oncogene Proteins c-akt; D000072377:Syk Kinase; D000099149:Ezrin; C000606896:SYK protein, human