The cytoplasmic domain mediates localization of furin to the trans-Golgi network en route to the endosomal/lysosomal system.
To investigate the mechanisms of membrane protein localization to the Golgi complex, we have examined the intracellular trafficking of epitope-tagged forms of the mammalian endopeptidase, furin, in stably transformed rat basophilic leukemia cells. Our studies show that furin is predominantly localized to the trans-Golgi network (TGN) at steady state, with smaller amounts present in intracellular vesicles. Biochemical and morphological analyses reveal that furin is progressively delivered to a lysosomal compartment, where it is degraded. Analyses of furin deletion mutants and chimeric proteins show that the cytoplasmic domain is both necessary and sufficient for localization to the TGN in various cell types. Interestingly, deletion of most of the cytoplasmic domain of furin results in a molecule that is predominantly localized to intracellular vesicles, some of which display characteristics of lysosomes. To a lesser extent, the cytoplasmically deleted molecule is also localized to the plasma membrane. These observations suggest the existence of an additional determinant for targeting to the endosomal/lysosomal system within the lumenal and/or transmembrane domains of furin. Thus, the overall pattern of trafficking and steady state localization of furin are determined by targeting information contained within more than one region of the molecule.
Bosshart
H
H
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Humphrey
J
J
Deignan
E
E
Davidson
J
J
Drazba
J
J
Yuan
L
L
Oorschot
V
V
Peters
P J
PJ
Bonifacino
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D002451:Cell Compartmentation; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D045683:Furin; D006056:Golgi Apparatus / Q000378:metabolism*; D007158:Immunologic Techniques; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D051381:Rats; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism*
D008562:Membrane Glycoproteins; D013381:Subtilisins; D045683:Furin