The membrane spanning domain of beta-1,4-galactosyltransferase specifies trans Golgi localization.
Chimeric cDNAs were constructed so as to generate hybrid proteins in which different parts of the N-terminal domain of the human invariant chain were replaced by equivalent sequences from the trans Golgi resident enzyme, beta-1,4-galactosyltransferase. The cytoplasmic and membrane spanning domains of galactosyltransferase were found to be sufficient to retain all of the hybrid invariant chain in trans Golgi cisternae as judged by indirect immunofluorescence, treatment with brefeldin A and immuno-electron microscopy. As few as ten amino acids corresponding to the lumenal half of the membrane spanning domain of the Golgi enzyme sufficed to localize most of the hybrid invariant chain to the trans cisternae. A cytoplasmic domain was necessary for complete retention as assessed by flow cytofluorometry but could be provided either by galactosyltransferase or by invariant chain. This suggests that the cytoplasmic domain plays a role accessory to the membrane spanning domain, the latter mediating compartmental specificity.
Cell Biology Laboratory, Imperial Cancer Research Fund, London, UK.
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Nilsson
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Lucocq
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Mackay
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Warren
D000595:Amino Acid Sequence; D000944:Antigens, Differentiation, B-Lymphocyte; D020126:Brefeldin A; D002462:Cell Membrane / Q000201:enzymology; D003517:Cyclopentanes / Q000494:pharmacology; D003593:Cytoplasm / Q000201:enzymology; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006056:Golgi Apparatus / Q000201:enzymology*; D006367:HeLa Cells / Q000648:ultrastructure; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D000944:Antigens, Differentiation, B-Lymphocyte; D003517:Cyclopentanes; D000949:Histocompatibility Antigens Class II; C045084:invariant chain; D020126:Brefeldin A; D004247:DNA; D009237:N-Acetyllactosamine Synthase