The Golgi matrix protein GM130: a specific interacting partner of the small GTPase rab1b.
To detect specific partners of the small Golgi-localized GTPase rab1b we generated rab1b mutants and used them as bait proteins in yeast two-hybrid screens. We isolated several specifically interacting clones. Two of them encode large protein fragments highly homologous to rat GM130 and to human Golgin95. The full-length human GM130 cDNA was cloned and its interaction with rab1b was characterized in detail by yeast two-hybrid and in vitro binding assays. Here we report for the first time that the rab1b protein interacts specifically with GM130 in a GTP-dependent manner and therefore needs the hypervariable regions of the N- and C-termini. We mapped the rab1b binding site of GM130 and provide evidence that it is different to the previously described p115 and Grasp65 binding sites of the GM130 protein.
Weide
T
T
Department of Experimental Tumorbiology, University of Muenster, Badestrasse 9, D-48149 Muenster, Germany.
Bayer
M
M
Köster
M
M
Siebrasse
J P
JP
Peters
R
R
Barnekow
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D006056:Golgi Apparatus / Q000378:metabolism*; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins; D001616:beta-Galactosidase / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D020761:rab1 GTP-Binding Proteins
D001324:Autoantigens; D002352:Carrier Proteins; D018076:DNA, Complementary; C108705:GORASP1 protein, human; D000077614:Golgi Matrix Proteins; C097741:Golgin subfamily A member 2; C491577:Gorasp1 protein, rat; D008565:Membrane Proteins; D011994:Recombinant Proteins; D033921:Vesicular Transport Proteins; C091580:vesicular transport factor p115; D001616:beta-Galactosidase; C061569:RAB1B protein, rat; D020691:rab GTP-Binding Proteins; D020761:rab1 GTP-Binding Proteins