The cytoplasmic tail of alpha 1,2-fucosyltransferase contains a sequence for golgi localization.
The Golgi apparatus has a central role in the glycosylation of proteins and lipids. There is a sequential addition of carbohydrates by glycosyltransferases that are distributed within the Golgi in the order in which the glycosylation occurs. The mechanism of glycosyltransferase retention is considered to involve their transmembrane domains and flanking regions, although we have shown that the cytoplasmic tail of alpha1,2-fucosyltransferase is important for its Golgi localization. Here we show that the removal of the alpha1,2-fucosyltransferase cytoplasmic tail altered its function of fucosylation and its localization site. When the tail was removed, the enzyme moved from the Golgi to the trans Golgi network, suggesting that the transmembrane is responsible for retention and that the cytoplasmic tail is responsible for localization. The cytoplasmic tail of alpha1,2-fucosyltransferase contains 8 amino acids (MWVPSRRH), and mutating these to alanine indicated a role for amino acids 3 to 7 in localization with a particular role of Ser(5). Mutagenesis of Ser(5) to amino acids containing an hydroxyl (Tyr and Thr) demonstrated that the hydroxyl at position 5 is important. Thus, the cytoplasmic tail, and especially a single amino acid, has a predominant role in the localization and thus the function of alpha1,2-fucosyltransferase.
Milland
J
J
John Connell Laboratory for Glycobiology, The Austin Research Institute, Studley Road, Heidelberg 3084, Australia.
Taylor
S G
SG
Dodson
H C
HC
McKenzie
I F
IF
Sandrin
M S
MS
D000595:Amino Acid Sequence; D001483:Base Sequence; D003593:Cytoplasm / Q000201:enzymology*; D017931:DNA Primers; D005647:Fucosyltransferases / Q000737:chemistry / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology*; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D017931:DNA Primers; D005647:Fucosyltransferases; D000097631:Galactoside 2-alpha-L-fucosyltransferase