title: G1 Domain of Versican Regulates Hyaluronan Organization and the Phenotype of Cultured Human Dermal Fibroblasts.
abstract: Variants of versican have wide-ranging effects on cell and tissue phenotype, impacting proliferation, adhesion, pericellular matrix composition, and elastogenesis. The G1 domain of versican, which contains two Link modules that bind to hyaluronan (HA), may be central to these effects. Recombinant human G1 (rhG1) with an N-terminal 8 amino acid histidine (His) tag, produced in Nicotiana benthamiana, was applied to cultures of dermal fibroblasts, and effects on proliferation and pericellular HA organization determined. rhG1 located to individual strands of cell surface HA which aggregated into structures resembling HA cables. On both individual and aggregated strands, the spacing of attached rhG1 was similar (~120 nm), suggesting interaction between rhG1 molecules. Endogenous V0/V1, present on HA between attached rhG1, did not prevent cable formation, while treatment with V0/V1 alone, which also bound to HA, did not induce cables. A single treatment with rhG1 suppressed cell proliferation for an extended period. Treating cells for 4 weeks with rhG1 resulted in condensed layers of elongated, differentiated α actin-positive fibroblasts, with rhG1 localized to cell surfaces, and a compact extracellular matrix including both collagen and elastin. These results demonstrate that the G1 domain of versican can regulate the organization of pericellular HA and affect phenotype.
authors.affiliation: Department of Anatomy and Medical Imaging, School of Medical Sciences, University of Auckland, Auckland, New Zealand (MJM,NZ)
authors.forename: Mervyn J
authors.initials: MJ
authors.lastname: Merrilees
authors.affiliation: Department of Anatomy and Medical Imaging, School of Medical Sciences, University of Auckland, Auckland, New Zealand (MJM,NZ)
authors.forename: Ning
authors.initials: N
authors.lastname: Zuo
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, Seattle, Washington (SPE,TNW)
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Evanko
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, The Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom (AJD)
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Day
authors.affiliation: Matrix Biology Program, Benaroya Research Institute, Seattle, Washington (SPE,TNW)
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Wight
mesh_terms: D049109:Cell Proliferation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism*; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000166:cytology*; D053675:Versicans / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; C508322:VCAN protein, human; D053675:Versicans; D006639:Histidine; D006820:Hyaluronic Acid
keywords: G1 domain; cables; differentiation; hyaluronan; versican