title: Polyphosphoinositides-dependent regulation of the osteoclast actin cytoskeleton and bone resorption.
abstract: BACKGROUND
  Gelsolin, an actin capping protein of osteoclast podosomes, has a unique function in regulating assembly and disassembly of the podosome actin filament. Previously, we have reported that osteopontin (OPN) binding to integrin alphavbeta3 increased the levels of gelsolin-associated polyphosphoinositides, podosome assembly/disassembly, and actin filament formation. The present study was undertaken to identify the possible role of polyphosphoinositides and phosphoinositides binding domains (PBDs) of gelsolin in the osteoclast cytoskeletal structural organization and osteoclast function.
  
  
  RESULTS
  Transduction of TAT/full-length gelsolin and PBDs containing gelsolin peptides into osteoclasts demonstrated: 1) F-actin enriched patches; 2) disruption of actin ring; 3) an increase in the association polyphosphoinositides (PPIs) with the transduced peptides containing PBDs. The above-mentioned effects were more pronounced with gelsolin peptide containing 2 tandem repeats of PBDs (PBD (2)). Binding of PPIs to the transduced peptides has resulted in reduced levels of PPIs association with the endogenous gelsolin, and thereby disrupted the actin remodeling processes in terms of podosome organization in the clear zone area and actin ring formation. These peptides also exhibited a dominant negative effect in the formation of WASP-Arp2/3 complex indicating the role of phosphoinositides in WASP activation. The TAT-PBD gelsolin peptides transduced osteoclasts are functionally defective in terms of motility and bone resorption.
  
  
  CONCLUSIONS
  Taken together, these data demonstrate that transduction of PBD gelsolin peptides into osteoclasts produced a dominant negative effect on actin assembly, motility, and bone resorption. These findings indicate that phosphoinositide-mediated signaling mechanisms regulate osteoclast cytoskeleton, podosome assembly/disassembly, actin ring formation and bone resorption activity of osteoclasts.
authors.affiliation: Department of Biomedical Sciences, University of Maryland, Baltimore, Maryland 21201, USA. mac001@dental.umaryland.edu
authors.forename: Rajat S
authors.initials: RS
authors.lastname: Biswas
authors.forename: DeAnna
authors.initials: D
authors.lastname: Baker
authors.forename: Keith A
authors.initials: KA
authors.lastname: Hruska
authors.forename: Meenakshi A
authors.initials: MA
authors.lastname: Chellaiah
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure*; D000199:Actins / Q000032:analysis; D000818:Animals; D001862:Bone Resorption; D018260:Gelsolin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D010010:Osteoclasts / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure*; D053495:Osteopontin; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012795:Sialoglycoproteins / Q000494:pharmacology; D014161:Transduction, Genetic
chemical_list: D000199:Actins; D018260:Gelsolin; D008840:Microfilament Proteins; D010455:Peptides; D018129:Phosphatidylinositol Phosphates; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D053495:Osteopontin; D019869:Phosphatidylinositol 3-Kinases