title: The formin FRL1 (FMNL1) is an essential component of macrophage podosomes.
abstract: Podosomes are highly dynamic actin-rich adhesion structures in cells of myeloid lineage and some transformed cells. Unlike transformed mesenchymal cell types, podosomes are the sole adhesion structure in macrophage and thus mediate all contact with adhesion substrate, including movement through complex tissues for immune surveillance. The existence of podosomes in inflammatory macrophages and transformed cell types suggest an important role in tissue invasion. The proteome, assembly, and maintenance of podosomes are emerging, but remain incompletely defined. Previously, we reported a formin homology sequence and actin assembly activity in association with macrophage beta-3 integrin. In this study we demonstrate by quantitative reverse transcriptase polymerase chain reaction and Western blotting that the formin FRL1 is specifically upregulated during monocyte differentiation to macrophages. We show that the formin FRL1 localizes to the actin-rich cores of primary macrophage podosomes. FRL1 co-precipitates with beta-3 integrin and both fixed and live cell fluorescence microscopy show that endogenous and overexpressed FRL1 selectively localize to macrophage podosomes. Targeted disruption of FRL1 by siRNA results in reduced cell adhesion and disruption of podosome dynamics. Our data suggest that FRL1 is responsible for modifying actin at the macrophage podosome and may be involved in actin cytoskeleton dynamics during adhesion and migration within tissues.
authors.affiliation: Department of Cell and Development Biology, SUNY Upstate Medical University, Syracuse, New York 13210, USA.
authors.forename: Akos T
authors.initials: AT
authors.lastname: Mersich
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Miller
authors.forename: Halina
authors.initials: H
authors.lastname: Chkourko
authors.forename: Scott D
authors.initials: SD
authors.lastname: Blystone
mesh_terms: D008841:Actin Cytoskeleton / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D022081:Cell Surface Extensions / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000502:physiology*; D005326:Fetal Proteins / Q000502:physiology*; D000080865:Formins; D006801:Humans; D039661:Integrin beta3 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008264:Macrophages / Q000166:cytology*; D008840:Microfilament Proteins / Q000502:physiology*; D009687:Nuclear Proteins / Q000502:physiology*; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; C476204:FMNL1 protein, human; D005326:Fetal Proteins; D000080865:Formins; D039661:Integrin beta3; D008840:Microfilament Proteins; D009687:Nuclear Proteins; D020741:rho GTP-Binding Proteins