RGD Reference Report - In vivo dynamics of the F-actin-binding protein neurabin-II. - Rat Genome Database

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In vivo dynamics of the F-actin-binding protein neurabin-II.

Authors: Stephens, DJ  Banting, G 
Citation: Stephens DJ and Banting G, Biochem J. 2000 Jan 15;345 Pt 2:185-94.
RGD ID: 10002737
Pubmed: PMID:10620493   (View Abstract at PubMed)
PMCID: PMC1220745   (View Article at PubMed Central)

Neurabin-II (spinophilin) is a ubiquitously expressed F-actin-binding protein containing an N-terminal actin-binding domain, a PDZ (PSD95/discs large/ZO-1) domain and a C-terminal domain predicted to form a coiled-coil structure. We have stably expressed a green fluorescent protein (GFP)-tagged version of neurabin-II in PC12 cells, and characterized the in vivo dynamics of this actin-binding protein using confocal fluorescence microscopy. We show that GFP-neurabin-II localizes to actin filaments, especially at cortical sites and areas underlying sites of active membrane remodelling. GFP-neurabin-II labels only a subset of F-actin within these cells, as indicated by rhodamine-phalloidin staining. Both actin filaments and small, highly motile structures within the cell body are seen. Photobleaching experiments show that GFP-neurabin-II also exhibits highly dynamic behaviour when bound to actin filaments. Latrunculin B treatment results in rapid relocalization of GFP-neurabin-II to the cytosol, whereas cytochalasin D treatment causes the collapse of GFP-neurabin-II fluorescence to intensely fluorescent foci of F-actin within the cell body. This collapse is reversed on cytochalasin D removal, recovery from which is greatly accelerated by stimulation of cells with epidermal growth factor (EGF). Furthermore, we show that this EGF-induced relocalization of GFP-neurabin-II is dependent on the activity of the small GTPase Rac1 but not the activity of ADP-ribosylation factor 6.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Ppp1r9bRatactin filament depolymerization  IEP  RGD 
Ppp1r9bRatcellular response to epidermal growth factor stimulus  IEP  RGD 

Cellular Component

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Ppp1r9bRatneuronal cell body  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Ppp1r9b  (protein phosphatase 1, regulatory subunit 9B)


Additional Information