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EMBO J. Dec 1991; 10(12): 3567–3575.
PMCID: PMC453088

The membrane spanning domain of beta-1,4-galactosyltransferase specifies trans Golgi localization.

Abstract

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.

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  • Rose JK, Doms RW. Regulation of protein export from the endoplasmic reticulum. Annu Rev Cell Biol. 1988;4:257–288. [PubMed]
  • Sege K, Rask L, Peterson PA. Role of beta2-microglobulin in the intracellular processing of HLA antigens. Biochemistry. 1981 Aug 4;20(16):4523–4530. [PubMed]
  • Stirzaker SC, Both GW. The signal peptide of the rotavirus glycoprotein VP7 is essential for its retention in the ER as an integral membrane protein. Cell. 1989 Mar 10;56(5):741–747. [PubMed]
  • Strous GJ. Golgi and secreted galactosyltransferase. CRC Crit Rev Biochem. 1986;21(2):119–151. [PubMed]
  • Strubin M, Mach B, Long EO. The complete sequence of the mRNA for the HLA-DR-associated invariant chain reveals a polypeptide with an unusual transmembrane polarity. EMBO J. 1984 Apr;3(4):869–872. [PMC free article] [PubMed]
  • Szczesna-Skorupa E, Kemper B. NH2-terminal substitutions of basic amino acids induce translocation across the microsomal membrane and glycosylation of rabbit cytochrome P450IIC2. J Cell Biol. 1989 Apr;108(4):1237–1243. [PMC free article] [PubMed]
  • von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. [PubMed]
  • Warren G. Protein transport. Signals and salvage sequences. Nature. 1987 May 7;327(6117):17–18. [PubMed]
  • Weinstein J, Lee EU, McEntee K, Lai PH, Paulson JC. Primary structure of beta-galactoside alpha 2,6-sialyltransferase. Conversion of membrane-bound enzyme to soluble forms by cleavage of the NH2-terminal signal anchor. J Biol Chem. 1987 Dec 25;262(36):17735–17743. [PubMed]
  • Wickner WT, Lodish HF. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. [PubMed]
  • Wiedmann M, Kurzchalia TV, Hartmann E, Rapoport TA. A signal sequence receptor in the endoplasmic reticulum membrane. Nature. 328(6133):830–833. [PubMed]
  • Wraight CJ, van Endert P, Möller P, Lipp J, Ling NR, MacLennan IC, Koch N, Moldenhauer G. Human major histocompatibility complex class II invariant chain is expressed on the cell surface. J Biol Chem. 1990 Apr 5;265(10):5787–5792. [PubMed]
  • Armstrong J, Patel S. The Golgi sorting domain of coronavirus E1 protein. J Cell Sci. 1991 Apr;98(Pt 4):567–575. [PubMed]
  • Bakke O, Dobberstein B. MHC class II-associated invariant chain contains a sorting signal for endosomal compartments. Cell. 1990 Nov 16;63(4):707–716. [PubMed]
  • Blobel G, Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. [PMC free article] [PubMed]
  • Bormann BJ, Knowles WJ, Marchesi VT. Synthetic peptides mimic the assembly of transmembrane glycoproteins. J Biol Chem. 1989 Mar 5;264(7):4033–4037. [PubMed]
  • Cosson P, Lankford SP, Bonifacino JS, Klausner RD. Membrane protein association by potential intramembrane charge pairs. Nature. 1991 May 30;351(6325):414–416. [PubMed]
  • Doms RW, Russ G, Yewdell JW. Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum. J Cell Biol. 1989 Jul;109(1):61–72. [PMC free article] [PubMed]
  • Dunphy WG, Brands R, Rothman JE. Attachment of terminal N-acetylglucosamine to asparagine-linked oligosaccharides occurs in central cisternae of the Golgi stack. Cell. 1985 Feb;40(2):463–472. [PubMed]
  • Freedman RB. Protein disulfide isomerase: multiple roles in the modification of nascent secretory proteins. Cell. 1989 Jun 30;57(7):1069–1072. [PubMed]
  • Fujiwara T, Oda K, Yokota S, Takatsuki A, Ikehara Y. Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. J Biol Chem. 1988 Dec 5;263(34):18545–18552. [PubMed]
  • Griffiths G, McDowall A, Back R, Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. [PubMed]
  • Grundmann U, Nerlich C, Rein T, Zettlmeissl G. Complete cDNA sequence encoding human beta-galactoside alpha-2,6-sialyltransferase. Nucleic Acids Res. 1990 Feb 11;18(3):667–667. [PMC free article] [PubMed]
  • Görlich D, Prehn S, Hartmann E, Herz J, Otto A, Kraft R, Wiedmann M, Knespel S, Dobberstein B, Rapoport TA. The signal sequence receptor has a second subunit and is part of a translocation complex in the endoplasmic reticulum as probed by bifunctional reagents. J Cell Biol. 1990 Dec;111(6 Pt 1):2283–2294. [PMC free article] [PubMed]
  • Hurtley SM, Helenius A. Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:277–307. [PubMed]
  • Jackson MR, Nilsson T, Peterson PA. Identification of a consensus motif for retention of transmembrane proteins in the endoplasmic reticulum. EMBO J. 1990 Oct;9(10):3153–3162. [PMC free article] [PubMed]
  • Kuroki K, Russnak R, Ganem D. Novel N-terminal amino acid sequence required for retention of a hepatitis B virus glycoprotein in the endoplasmic reticulum. Mol Cell Biol. 1989 Oct;9(10):4459–4466. [PMC free article] [PubMed]
  • Lauffer L, Garcia PD, Harkins RN, Coussens L, Ullrich A, Walter P. Topology of signal recognition particle receptor in endoplasmic reticulum membrane. Nature. 318(6044):334–338. [PubMed]
  • Lipp J, Dobberstein B. The membrane-spanning segment of invariant chain (I gamma) contains a potentially cleavable signal sequence. Cell. 1986 Sep 26;46(7):1103–1112. [PubMed]
  • Lippincott-Schwartz J, Yuan LC, Bonifacino JS, Klausner RD. Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell. 1989 Mar 10;56(5):801–813. [PubMed]
  • Lotteau V, Teyton L, Peleraux A, Nilsson T, Karlsson L, Schmid SL, Quaranta V, Peterson PA. Intracellular transport of class II MHC molecules directed by invariant chain. Nature. 1990 Dec 13;348(6302):600–605. [PubMed]
  • Louvard D, Reggio H, Warren G. Antibodies to the Golgi complex and the rough endoplasmic reticulum. J Cell Biol. 1982 Jan;92(1):92–107. [PMC free article] [PubMed]
  • Lucocq JM, Pryde JG, Berger EG, Warren G. A mitotic form of the Golgi apparatus in HeLa cells. J Cell Biol. 1987 Apr;104(4):865–874. [PMC free article] [PubMed]
  • Machamer CE, Rose JK. A specific transmembrane domain of a coronavirus E1 glycoprotein is required for its retention in the Golgi region. J Cell Biol. 1987 Sep;105(3):1205–1214. [PMC free article] [PubMed]
  • Machamer CE, Mentone SA, Rose JK, Farquhar MG. The E1 glycoprotein of an avian coronavirus is targeted to the cis Golgi complex. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6944–6948. [PMC free article] [PubMed]
  • Masri KA, Appert HE, Fukuda MN. Identification of the full-length coding sequence for human galactosyltransferase (beta-N-acetylglucosaminide: beta 1,4-galactosyltransferase). Biochem Biophys Res Commun. 1988 Dec 15;157(2):657–663. [PubMed]
  • Munro S, Pelham HR. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. [PubMed]
  • Nilsson I, von Heijne G. Fine-tuning the topology of a polytopic membrane protein: role of positively and negatively charged amino acids. Cell. 1990 Sep 21;62(6):1135–1141. [PubMed]
  • Nilsson T, Jackson M, Peterson PA. Short cytoplasmic sequences serve as retention signals for transmembrane proteins in the endoplasmic reticulum. Cell. 1989 Aug 25;58(4):707–718. [PubMed]
  • Omary MB, Trowbridge IS. Biosynthesis of the human transferrin receptor in cultured cells. J Biol Chem. 1981 Dec 25;256(24):12888–12892. [PubMed]
  • Päbo S, Bhat BM, Wold WS, Peterson PA. A short sequence in the COOH-terminus makes an adenovirus membrane glycoprotein a resident of the endoplasmic reticulum. Cell. 1987 Jul 17;50(2):311–317. [PubMed]
  • Parks GD, Lamb RA. Topology of eukaryotic type II membrane proteins: importance of N-terminal positively charged residues flanking the hydrophobic domain. Cell. 1991 Feb 22;64(4):777–787. [PubMed]
  • Paulson JC, Colley KJ. Glycosyltransferases. Structure, localization, and control of cell type-specific glycosylation. J Biol Chem. 1989 Oct 25;264(30):17615–17618. [PubMed]
  • Pelham HR. Control of protein exit from the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:1–23. [PubMed]
  • Pfeffer SR, Rothman JE. Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi. Annu Rev Biochem. 1987;56:829–852. [PubMed]

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