Inactive and active mutants of rab1b are not tightly integrated into target membranes.
The rab1b GTPase is localized on the ER and cis-Golgi membranes and involved in early exocytotic membrane traffic processes. To analyze the influence of rab1b conformation on specific membrane targeting processes three point mutants simulating permanently inactive or active rab1b proteins were constructed and expressed in BHK cells. To increase the expression of the growth inhibiting rab1b S22N and rab1b N121I mutants we co-expressed the Mss4 protein and observed an elevated expression of the inactive rab1b mutants. Surprisingly, only the rab1b wild-type protein shows the correct intracellular localization, and a tight membrane association. We conclude that the targeting process of rab1b depends predominantly on GDP/GTP exchange.
Weide
T
T
Department of Experimental Tumorbiology, University of Muenster, 48149 Muenster, Germany.
Koster
M
M
Barnekow
A
A
D019943:Amino Acid Substitution; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011506:Proteins / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D020691:rab GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D020761:rab1 GTP-Binding Proteins
D000939:Epitopes; D006389:Hemagglutinins, Viral; D011506:Proteins; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins; C061569:RAB1B protein, rat; D020691:rab GTP-Binding Proteins; D020761:rab1 GTP-Binding Proteins