Specific sequences in the signal anchor of the beta-galactoside alpha-2,6-sialyltransferase are not essential for Golgi localization. Membrane flanking sequences may specify Golgi retention.
The beta-galactoside alpha-2,6-sialyltransferase is a trans Golgi/trans Golgi network glycosyltransferase which adds sialic acid residues to Asn-linked oligosaccharides of glycoproteins. Previous results suggested that the sialyltransferase stem and signal anchor including flanking sequences may be two independent Golgi retention regions. However, other experiments demonstrated that the sequence of the signal anchor itself was not important. To investigate whether the sialyltransferase signal anchor was necessary and sufficient for Golgi retention, several mutant and chimeric proteins were expressed and localized in Cos-1 and Chinese hamster ovary cells. We found that the signal anchor and flanking sequences were able to retain the sialyltransferase catalytic domain in the Golgi. However, efficient Golgi retention was still observed when the signal anchor was altered or entirely replaced in either the presence or absence of most of the luminal stem region. Chimeric proteins consisting of the sialyltransferase cytoplasmic tail and signal anchor fused to the extracellular domains of two different cell surface proteins demonstrated poor Golgi retention. A significant increase in the Golgi retention of one of these chimeras was observed when two lysines were placed next to the signal anchor on the luminal side. Taken together these results suggest that the sialyltransferase signal anchor is not necessary or sufficient for Golgi retention, rather, appropriately spaced cytoplasmic and luminal flanking sequences are the important elements of the sialyltransferase Golgi retention region.
Dahdal
R Y
RY
Department of Biochemistry, University of Illinois at Chicago, College of Medicine 60612.
Colley
K J
KJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002384:Catalysis; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers; D006056:Golgi Apparatus / Q000201:enzymology*; D007425:Intracellular Membranes / Q000737:chemistry; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000378:metabolism; D021382:Protein Sorting Signals / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012799:Sialyltransferases / Q000737:chemistry* / Q000378:metabolism; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D017931:DNA Primers; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D012799:Sialyltransferases; D009439:Neuraminidase; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase