title: Myosin IIA Heavy Chain Phosphorylation Mediates Adhesion Maturation and Protrusion in Three Dimensions.
abstract: Non-muscle myosin II (NMII) is a conserved force-producing cytoskeletal enzyme with important but poorly understood roles in cell migration. To investigate myosin heavy chain (MHC) phosphorylation roles in 3D migration, we expressed GFP-tagged NMIIA wild-type or mutant constructs in cells depleted of endogenous NMIIA protein. We find that individual mutation or double mutation of Ser-1916 or Ser-1943 to alanine potently blocks recruitment of GFP-NM-IIA filaments to leading edge protrusions in 2D, and this in turn blocks maturation of anterior focal adhesions. When placed in 3D collagen gels, cells expressing wild-type GFP MHC-IIA behave like parental cells, displaying robust and active formation and retraction of protrusions. However, cells depleted of NMIIA or cells expressing the mutant GFP MHC-IIA display severe defects in invasion and in stabilizing protrusions in 3D. These studies reveal an NMIIA-specific role in 3D invasion that requires competence for NMIIA phosphorylation at Ser-1916 and Ser-1943. In sum, these results demonstrate a critical and previously unrecognized role for NMIIA phosphorylation in 3D invasion.
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Vandana
authors.initials: V
authors.lastname: Rai
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Dustin G
authors.initials: DG
authors.lastname: Thomas
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Jordan R
authors.initials: JR
authors.lastname: Beach
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195. Electronic address: tte@case.edu.
authors.forename: Thomas T
authors.initials: TT
authors.lastname: Egelhoff
mesh_terms: D000818:Animals; D019556:COS Cells; D002448:Cell Adhesion; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement; D002522:Chlorocebus aethiops; D006801:Humans; D051379:Mice; D018995:Myosin Heavy Chains / Q000032:analysis / Q000378:metabolism*; D024507:Nonmuscle Myosin Type IIA / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation
chemical_list: D024507:Nonmuscle Myosin Type IIA; D018995:Myosin Heavy Chains
keywords: cell migration; cytoskeleton; extracellular matrix; invasion; myosin