A novel partner for Dictyostelium filamin is an alpha-helical developmentally regulated protein

Journal of Cell Science
Monika KnuthAngelika A Noegel

Abstract

The filamins are a family of highly homologous actin-crosslinking proteins that stabilize three-dimensional actin networks, link them to membrane proteins and direct intracellular signaling reactions to the actin scaffold through interaction with various binding partners. Here, we describe the first Dictyostelium filamin-interacting protein to be isolated--FIP, a 229.8 kDa protein with two alpha-helical coiled coil domains. FIP was identified in a yeast two-hybrid screen using the rod domain of filamin as bait. FIP can also be coimmunoprecipitated with filamin from cellular extracts. Deletion analysis located the interaction domain of FIP to a C-terminal region; by contrast, in filamin rods, repeats 2-4 interacted with the recombinant FIP protein. The 7 kb transcript of FIP is upregulated during early development. Monoclonal antibodies raised against a bacterially expressed FIP polypeptide recognize a 230 kDa developmentally regulated protein in western blots. Immunofluorescence analysis shows a punctate staining pattern in the cytosol and, in cell fractionation experiments, FIP is mainly found in the cytosolic fraction. A fusion protein composed of GFP and the C-terminal part localizes to the plasma membrane and is associated ...Continue Reading

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Citations

May 10, 2011·Molecular Biology of the Cell·Sebastian KickaRichard A Firtel
Nov 14, 2007·BMC Cell Biology·Sarah J AnnesleyPaul R Fisher
Aug 10, 2005·Genome Biology·Andrew WilkinsRobert H Insall
Feb 27, 2010·PloS One·Maria Christina ShinaWaldemar Kolanus
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Dec 7, 2010·Seminars in Cell & Developmental Biology·Lisa M FrancionePaul R Fisher
Nov 13, 2009·Current Biology : CB·André E X Brown, Dennis E Discher

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