title: Adapter protein SH2B1beta cross-links actin filaments and regulates actin cytoskeleton.
abstract: The Src homology 2 (SH2) domain-containing adapter protein SH2B1beta plays a role in severe obesity, leptin and insulin resistance, and infertility. SH2B1beta was initially identified as a Janus tyrosine kinase 2 (JAK2) substrate, and it has been implicated in cell motility and regulation of the actin rearrangement in response to GH and platelet-derived growth factor. SH2B1beta is also required for maximal actin-based motility of Listeria. Here we have used a low-speed pelleting assay and electron microscopy to demonstrate that SH2B1beta has two actin-binding sites and that it cross-links actin filaments in vitro. Wild-type SH2B1beta localized to cell ruffles and along filopodia, but deletion of amino acids 150-200 (the first actin-binding site) led to mislocalization of the protein to filopodia tip complexes where it colocalized with vasodilator-stimulated phosphoprotein (VASP). Based on studies performed in VASP-deficient MVD7(-/-) cells, with or without green fluorescent protein-VASP reconstitution, we concluded that the proper intracellular localization of native SH2B1beta required the presence of the first SH2B1beta actin-binding site and VASP. Finally, we found that both SH2B1beta actin-binding domains were required for maximal GH- and prolactin-induced cell ruffling. Together, these results suggest that SH2B1beta functions as an adapter protein that cross-links actin filaments, leading to modulation of cellular responses in response to JAK2 activation.
authors.affiliation: Department of Biological Sciences, University of Toledo, Toledo, Ohio 43606-3390, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Rider
authors.forename: Jing
authors.initials: J
authors.lastname: Tao
authors.forename: Stacy
authors.initials: S
authors.lastname: Snyder
authors.forename: Brittany
authors.initials: B
authors.lastname: Brinley
authors.forename: Jiayun
authors.initials: J
authors.lastname: Lu
authors.forename: Maria
authors.initials: M
authors.lastname: Diakonova