Rab22a regulates the recycling of membrane proteins internalized independently of clathrin.
Plasma membrane proteins that are internalized independently of clathrin, such as major histocompatibility complex class I (MHCI), are internalized in vesicles that fuse with the early endosomes containing clathrin-derived cargo. From there, MHCI is either transported to the late endosome for degradation or is recycled back to the plasma membrane via tubular structures that lack clathrin-dependent recycling cargo, e.g., transferrin. Here, we show that the small GTPase Rab22a is associated with these tubular recycling intermediates containing MHCI. Expression of a dominant negative mutant of Rab22a or small interfering RNA-mediated depletion of Rab22a inhibited both formation of the recycling tubules and MHCI recycling. By contrast, cells expressing the constitutively active mutant of Rab22a exhibited prominent recycling tubules and accumulated vesicles at the periphery, but MHCI recycling was still blocked. These results suggest that Rab22a activation is required for tubule formation and Rab22a inactivation for final fusion of recycling membranes with the surface. The trafficking of transferrin was only modestly affected by these treatments. Dominant negative mutant of Rab11a also inhibited recycling of MHCI but not the formation of recycling tubules, suggesting that Rab22a and Rab11a might coordinate different steps of MHCI recycling.
Weigert
Roberto
R
Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-8017, USA.
Yeung
Albert Chi
AC
Li
Jean
J
Donaldson
Julie G
JG
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000502:physiology; D002966:Clathrin / Q000378:metabolism*; D011992:Endosomes / Q000276:immunology / Q000502:physiology; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D021381:Protein Transport / Q000502:physiology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D011990:Receptors, Transferrin / Q000235:genetics / Q000502:physiology; D014168:Transferrin / Q000378:metabolism; D014617:Vacuoles / Q000276:immunology / Q000502:physiology; D020691:rab GTP-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D000099193:rab11 GTP-Binding Proteins
D002966:Clathrin; D015395:Histocompatibility Antigens Class I; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D011990:Receptors, Transferrin; D014168:Transferrin; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C484191:RAB22A protein, human