Localization of furin to the trans-Golgi network and recycling from the cell surface involves Ser and Tyr residues within the cytoplasmic domain.
Furin is a membrane-associated endoprotease that catalyzes cleavage of precursor proteins at Arg-X-Lys/Arg-Arg sites. Although, at steady state, furin is predominantly found in the trans-Golgi network (TGN), it also cycles between the TGN and the cell surface. Recently, the cytoplasmic tail of furin has been shown to be sufficient for its localization to the TGN. Within the cytoplasmic domain, there are Ser residues, which we now show are sites for phosphorylation by casein kinase II in vitro, and a Tyr-containing sequence, both of which have been shown to be important for other TGN proteins to localize to this compartment. In the present study, we show by site-directed mutagenesis that these residues are important for TGN localization and recycling of furin. Mutation of the Ser residues abrogated the TGN localization. By contrast, mutation of the Tyr residue did not affect the TGN localization but impaired the internalization from the plasma membrane. These observations suggest that distinct cytoplasmic determinants are responsible for retention in the TGN and retrieval from the cell surface of furin.
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D015704:CD4 Antigens / Q000096:biosynthesis; D047390:Casein Kinase II; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000201:enzymology; D045683:Furin; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D007668:Kidney; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D012694:Serine; D013379:Substrate Specificity; D013381:Subtilisins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection; D014443:Tyrosine
D015703:Antigens, CD; D015704:CD4 Antigens; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014443:Tyrosine; D012694:Serine; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D013381:Subtilisins; D045683:Furin