title: Wdpcp, a PCP protein required for ciliogenesis, regulates directional cell migration and cell polarity by direct modulation of the actin cytoskeleton.
abstract: Planar cell polarity (PCP) regulates cell alignment required for collective cell movement during embryonic development. This requires PCP/PCP effector proteins, some of which also play essential roles in ciliogenesis, highlighting the long-standing question of the role of the cilium in PCP. Wdpcp, a PCP effector, was recently shown to regulate both ciliogenesis and collective cell movement, but the underlying mechanism is unknown. Here we show Wdpcp can regulate PCP by direct modulation of the actin cytoskeleton. These studies were made possible by recovery of a Wdpcp mutant mouse model. Wdpcp-deficient mice exhibit phenotypes reminiscent of Bardet-Biedl/Meckel-Gruber ciliopathy syndromes, including cardiac outflow tract and cochlea defects associated with PCP perturbation. We observed Wdpcp is localized to the transition zone, and in Wdpcp-deficient cells, Sept2, Nphp1, and Mks1 were lost from the transition zone, indicating Wdpcp is required for recruitment of proteins essential for ciliogenesis. Wdpcp is also found in the cytoplasm, where it is localized in the actin cytoskeleton and in focal adhesions. Wdpcp interacts with Sept2 and is colocalized with Sept2 in actin filaments, but in Wdpcp-deficient cells, Sept2 was lost from the actin cytoskeleton, suggesting Wdpcp is required for Sept2 recruitment to actin filaments. Significantly, organization of the actin filaments and focal contacts were markedly changed in Wdpcp-deficient cells. This was associated with decreased membrane ruffling, failure to establish cell polarity, and loss of directional cell migration. These results suggest the PCP defects in Wdpcp mutants are not caused by loss of cilia, but by direct disruption of the actin cytoskeleton. Consistent with this, Wdpcp mutant cochlea has normal kinocilia and yet exhibits PCP defects. Together, these findings provide the first evidence, to our knowledge, that a PCP component required for ciliogenesis can directly modulate the actin cytoskeleton to regulate cell polarity and directional cell migration.
authors.affiliation: Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America ; Laboratory of Developmental Biology, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Cheng
authors.initials: C
authors.lastname: Cui
authors.forename: Bishwanath
authors.initials: B
authors.lastname: Chatterjee
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Lozito
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Francis
authors.forename: Hisato
authors.initials: H
authors.lastname: Yagi
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Swanhart
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Sanker
authors.forename: Deanne
authors.initials: D
authors.lastname: Francis
authors.forename: Qing
authors.initials: Q
authors.lastname: Yu
authors.forename: Jovenal T
authors.initials: JT
authors.lastname: San Agustin
authors.forename: Chandrakala
authors.initials: C
authors.lastname: Puligilla
authors.forename: Tania
authors.initials: T
authors.lastname: Chatterjee
authors.forename: Terry
authors.initials: T
authors.lastname: Tansey
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Liu
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Kelley
authors.forename: Elias T
authors.initials: ET
authors.lastname: Spiliotis
authors.forename: Adam V
authors.initials: AV
authors.lastname: Kwiatkowski
authors.forename: Rocky
authors.initials: R
authors.lastname: Tuan
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Pazour
authors.forename: Neil A
authors.initials: NA
authors.lastname: Hukriede
authors.forename: Cecilia W
authors.initials: CW
authors.lastname: Lo
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002465:Cell Movement; D016764:Cell Polarity; D002478:Cells, Cultured; D002923:Cilia / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000502:physiology*; D004252:DNA Mutational Analysis; D022001:Focal Adhesions / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D021381:Protein Transport; D058112:Septins / Q000378:metabolism; D059008:Time-Lapse Imaging; D060449:Wnt Signaling Pathway; D015027:Zebrafish; D008565:Membrane Proteins
chemical_list: D003598:Cytoskeletal Proteins; D058112:Septins; C000589231:Wdpcp protein, mouse; D008565:Membrane Proteins; C106863:Sept2 protein, mouse