RGD Reference Report - Differential dynamics of Rab3A and Rab27A on secretory granules. - Rat Genome Database

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Differential dynamics of Rab3A and Rab27A on secretory granules.

Authors: Handley, Mark T W  Haynes, Lee P  Burgoyne, Robert D 
Citation: Handley MT, etal., J Cell Sci. 2007 Mar 15;120(Pt 6):973-84. doi: 10.1242/jcs.03406. Epub 2007 Feb 20.
RGD ID: 14390061
Pubmed: PMID:17311845   (View Abstract at PubMed)
PMCID: PMC1855981   (View Article at PubMed Central)
DOI: DOI:10.1242/jcs.03406   (Journal Full-text)

We have assessed the dynamics of the association of Rab3A and Rab27A with secretory granules at various stages of their life in PC12 cells. Endogenous Rab3A colocalised with the secretory granule marker secretogranin II (SGII) and expressed EGFP-Rab3A and ECFP-Rab27A colocalised with one another. The extent of colocalisation between EGFP-Rab3A or EGFP-Rab27 and SGII increased after longer times post transfection suggesting that these Rab proteins are preferentially recruited to newly synthesised granules. Following the release of immature secretory granules from the trans-Golgi network, Rab3A and Rab27A became associated with the immature granules after a lag period of around 20 minutes. Rab dynamics on granules were analysed in fluorescence recovery after photobleaching (FRAP) experiments. The recovery profile of EGFP-Rab27A was comparable to that of ppANF-EGFP, whereas the recovery profile of EGFP-Rab3A was significantly faster, indicating that Rab3A but not Rab27A might be rapidly exchanged between granules and cytosol. Inhibition of heat-shock protein 90 with 10 muM geldanamycin did not affect the exchange process or regulated exocytosis. Rab dynamics during stimulation with 300 muM ATP were analysed in live cells. Loss of granular ppANF-EGFP fluorescence was seen at the cell periphery after stimulation but only limited changes in EGFP-Rab3A and EGFP-Rab27A fluorescence was observed, indicating that the Rab proteins do not immediately dissociate or disperse on stimulation. The data suggest potentially distinct roles for Rab3A and Rab27A and we suggest that the finding that young secretory granules have a higher capacity for binding Rab3A and Rab27A is functionally important for preferential exocytosis from these granules.



Gene Ontology Annotations    Click to see Annotation Detail View

Cellular Component

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Rab27aRatsecretory granule located_inIDA PMID:17311845UniProt 
Rab3aRatsecretory granule located_inIDA PMID:17311845UniProt 

Objects Annotated

Genes (Rattus norvegicus)
Rab27a  (RAB27A, member RAS oncogene family)
Rab3a  (RAB3A, member RAS oncogene family)


Additional Information