title: Motor protein Myo1c is a podocyte protein that facilitates the transport of slit diaphragm protein Neph1 to the podocyte membrane.
abstract: The podocyte proteins Neph1 and nephrin organize a signaling complex at the podocyte cell membrane that forms the structural framework for a functional glomerular filtration barrier. Mechanisms regulating the movement of these proteins to and from the membrane are currently unknown. This study identifies a novel interaction between Neph1 and the motor protein Myo1c, where Myo1c plays an active role in targeting Neph1 to the podocyte cell membrane. Using in vivo and in vitro experiments, we provide data supporting a direct interaction between Neph1 and Myo1c which is dynamic and actin dependent. Unlike wild-type Myo1c, the membrane localization of Neph1 was significantly reduced in podocytes expressing dominant negative Myo1c. In addition, Neph1 failed to localize at the podocyte cell membrane and cell junctions in Myo1c-depleted podocytes. We further demonstrate that similarly to Neph1, Myo1c also binds nephrin and reduces its localization at the podocyte cell membrane. A functional analysis of Myo1c knockdown cells showed defects in cell migration, as determined by a wound assay. In addition, the ability to form tight junctions was impaired in Myo1c knockdown cells, as determined by transepithelial electric resistance (TER) and bovine serum albumin (BSA) permeability assays. These results identify a novel Myo1c-dependent molecular mechanism that mediates the dynamic organization of Neph1 and nephrin at the slit diaphragm and is critical for podocyte function.
authors.affiliation: Department of Medicine, Renal Electrolyte and Hypertension Division, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: E
authors.initials: E
authors.lastname: Arif
authors.forename: M C
authors.initials: MC
authors.lastname: Wagner
authors.forename: D B
authors.initials: DB
authors.lastname: Johnstone
authors.forename: H N
authors.initials: HN
authors.lastname: Wong
authors.forename: B
authors.initials: B
authors.lastname: George
authors.forename: P A
authors.initials: PA
authors.lastname: Pruthi
authors.forename: M J
authors.initials: MJ
authors.lastname: Lazzara
authors.forename: D
authors.initials: D
authors.lastname: Nihalani
mesh_terms: D000199:Actins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement / Q000235:genetics; D017097:Electric Impedance; D055785:Gene Knockdown Techniques; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D024461:Myosin Type I / Q000235:genetics / Q000378:metabolism*; D050199:Podocytes / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D019108:Tight Junctions / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; C431137:KIRREL1 protein, human; D008565:Membrane Proteins; D024461:Myosin Type I; C460367:MYO1C protein, human