RGD Reference Report - Expression and localization of two low molecular weight GTP-binding proteins, Rab8 and Rab10, by epitope tag. - Rat Genome Database

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Expression and localization of two low molecular weight GTP-binding proteins, Rab8 and Rab10, by epitope tag.

Authors: Chen, YT  Holcomb, C  Moore, HP 
Citation: Chen YT, etal., Proc Natl Acad Sci U S A 1993 Jul 15;90(14):6508-12.
RGD ID: 734514
Pubmed: PMID:7688123   (View Abstract at PubMed)
PMCID: PMC46961   (View Article at PubMed Central)

Small GTP-binding proteins of the YPT/SEC4/Rab family have been shown to play an essential role in intracellular membrane trafficking. In mammals, Rab8 and Rab10 are the two small GTP-binding proteins identified so far that are closest to SEC4, an essential gene product involved in post-Golgi constitutive secretion in the yeast Saccharomyces cerevisiae. To study the localization of Rab proteins, we have expressed the cDNAs with an influenza virus hemagglutinin (HA) epitope tag at the N terminus. The feasibility of this method was tested by using yeast SEC4. HA-tagged SEC4 functionally complemented a temperature-sensitive sec4 mutant similarly to wild-type SEC4, indicating that the modified protein retained functional integrity. Monoclonal antibody 12CA5, raised against the HA tag, was used to determine the expression and localization of HA-tagged proteins after transfection. In stably transfected CHO and Swiss 3T3 cells, HA-tagged Rab8 was localized to the cell periphery, with the highest concentration in the ruffling areas. In contrast, epitope-tagged Rab10 expressed in CHO and BHK cells was concentrated on membranes in the perinuclear region. By light microscopy, the staining partially overlapped with that of a Golgi marker, beta-COP. Thus, despite the high degree homology of Rab8 and Rab10 (66% identity), the two proteins are localized to distinct cellular compartments. This approach should provide a general tool for the analyses of other members of the YPT/SEC4/rab gene family.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
cellular response to antibiotic  IEP 734514brefeldin ARGD 

Cellular Component
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
perinuclear region of cytoplasm  IDA 734514 RGD 
trans-Golgi network  IDA 734514 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Rab10  (RAB10, member RAS oncogene family)


Additional Information