Rab GDP dissociation inhibitor as a general regulator for the membrane association of rab proteins.
Rab proteins comprise a family of small GTPases that serve a regulatory role in membrane traffic. These proteins are in part cytosolic and in part associated with the membranes of specific exocytic and endocytic organelles. Smg p25A/rab3A GDI, a cytosolic protein which inhibits the dissociation of GDP from smg p25A/rab3A, Sec4p, and rab11, has also been found to prevent association of rab3A with the membrane. In this study, we have used Madin-Darby canine kidney cells permeabilized with the bacterial toxin streptolysin O to test the general activity of rab3A GDI in modulating the membrane association of various small GTP-binding proteins. Rab3A GDP dissociation inhibitor (GDI) removed from the membrane all rab proteins we have tested and inhibited the membrane binding of in vitro translated rab proteins. However, rab3A GDI had a limited effect on the membrane association of a mutant rab5 protein which contained a farnesylated cysteine motif. Finally, we found that, although rab3A GDI resides primarily in the cytosol, it is also associated with compartments of the exocytic and endocytic pathways. Since rab3A GDI can modulate the membrane association of various rab proteins, we propose to rename it rab GDI.
Ullrich
O
O
European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
Stenmark
H
H
Alexandrov
K
K
Huber
L A
LA
Kaibuchi
K
K
Sasaki
T
T
Takai
Y
Y
Zerial
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D004285:Dogs; D004926:Escherichia coli / Q000235:genetics; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006706:Homeostasis; D007668:Kidney; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013301:Streptolysins / Q000494:pharmacology; D013816:Thermodynamics; D014158:Transcription, Genetic; D020692:rab3 GTP-Binding Proteins
D001426:Bacterial Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D013301:Streptolysins; C025964:streptolysin O; D005982:Glutathione Transferase; D019204:GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins