Opposing motor activities of dynein and kinesin determine retention and transport of MHC class II-containing compartments.
MHC class II molecules exert their function at the cell surface by presenting to T cells antigenic fragments that are generated in the endosomal pathway. The class II molecules are targetted to early lysosomal structures, termed MIIC, where they interact with antigenic fragments and are subsequently transported to the cell surface. We previously visualised vesicular transport of MHC class II-containing early lysosomes from the microtubule organising centre (MTOC) region towards the cell surface in living cells. Here we show that the MIIC move bidirectionally in a 'stop-and-go' fashion. Overexpression of a motor head-deleted kinesin inhibited MIIC motility, showing that kinesin is the motor that drives its plus end transport towards the cell periphery. Cytoplasmic dynein mediates the return of vesicles to the MTOC area and effectively retains the vesicles at this location, as assessed by inactivation of dynein by overexpression of dynamitin. Our data suggest a retention mechanism that determines the perinuclear accumulation of MIIC, which is the result of dynein activity being superior over kinesin activity. The bidirectional nature of MIIC movement is the result of both kinesin and dynein acting reciprocally on the MIIC during its transport. The motors may be the ultimate targets of regulatory kinases since the protein kinase inhibitor staurosporine induces a massive release of lysosomal vesicles from the MTOC region that is morphologically similar to that observed after inactivation of the dynein motor.
Netherlands Cancer Institute, Department of Tumor Biology, Plesmanlaan 121, The Netherlands.
R
R
Wubbolts
M
M
Fernandez-Borja
I
I
Jordens
E
E
Reits
S
S
Dusseljee
C
C
Echeverri
R B
RB
Vallee
J
J
Neefjes
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004398:Dyneins / Q000502:physiology*; D006681:HLA-D Antigens / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016547:Kinesins / Q000502:physiology*; D008247:Lysosomes / Q000502:physiology*; D018613:Microscopy, Confocal; D008870:Microtubules / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D011993:Recombinant Fusion Proteins; D004398:Dyneins; D016547:Kinesins