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. 1991 Dec 2;115(6):1521–1534. doi: 10.1083/jcb.115.6.1521

Isolation, characterization, and expression of cDNAs encoding murine alpha-mannosidase II, a Golgi enzyme that controls conversion of high mannose to complex N-glycans

PMCID: PMC2289207  PMID: 1757461

Abstract

Golgi alpha-mannosidase II (GlcNAc transferase I-dependent alpha 1,3[alpha 1,6] mannosidase, EC 3.2.1.114) catalyzes the final hydrolytic step in the N-glycan maturation pathway acting as the committed step in the conversion of high mannose to complex type structures. We have isolated overlapping clones from a murine cDNA library encoding the full length alpha-mannosidase II open reading frame and most of the 5' and 3' untranslated region. The coding sequence predicts a type II transmembrane protein with a short cytoplasmic tail (five amino acids), a single transmembrane domain (21 amino acids), and a large COOH-terminal catalytic domain (1,124 amino acids). This domain organization which is shared with the Golgi glycosyl-transferases suggests that the common structural motifs may have a functional role in Golgi enzyme function or localization. Three sets of polyadenylated clones were isolated extending 3' beyond the open reading frame by as much as 2,543 bp. Northern blots suggest that these polyadenylated clones totaling 6.1 kb in length correspond to minor message species smaller than the full length message. The largest and predominant message on Northern blots (7.5 kb) presumably extends another approximately 1.4-kb downstream beyond the longest of the isolated clones. Transient expression of the alpha-mannosidase II cDNA in COS cells resulted in 8-12-fold overexpression of enzyme activity, and the appearance of cross-reactive material in a perinuclear membrane array consistent with a Golgi localization. A region within the catalytic domain of the alpha-mannosidase II open reading frame bears a strong similarity to a corresponding sequence in the rat liver endoplasmic reticulum alpha-mannosidase and the vacuolar alpha- mannosidase of Saccharomyces cerevisiae. Partial human alpha- mannosidase II cDNA clones were also isolated and the gene was localized to human chromosome 5.

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Selected References

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  1. Appert H. E., Rutherford T. J., Tarr G. E., Wiest J. S., Thomford N. R., McCorquodale D. J. Isolation of a cDNA coding for human galactosyltransferase. Biochem Biophys Res Commun. 1986 Aug 29;139(1):163–168. doi: 10.1016/s0006-291x(86)80094-8. [DOI] [PubMed] [Google Scholar]
  2. Bischoff J., Moremen K., Lodish H. F. Isolation, characterization, and expression of cDNA encoding a rat liver endoplasmic reticulum alpha-mannosidase. J Biol Chem. 1990 Oct 5;265(28):17110–17117. [PubMed] [Google Scholar]
  3. Brandley B. K., Swiedler S. J., Robbins P. W. Carbohydrate ligands of the LEC cell adhesion molecules. Cell. 1990 Nov 30;63(5):861–863. doi: 10.1016/0092-8674(90)90487-y. [DOI] [PubMed] [Google Scholar]
  4. Carlock L. R., Smith D., Wasmuth J. J. Genetic counterselective procedure to isolate interspecific cell hybrids containing single human chromosomes: construction of cell hybrids and recombinant DNA libraries specific for human chromosomes 3 and 4. Somat Cell Mol Genet. 1986 Mar;12(2):163–174. doi: 10.1007/BF01560663. [DOI] [PubMed] [Google Scholar]
  5. Cheng S. H., Malcolm S., Pemble S., Winchester B. Purification and comparison of the structures of human liver acidic alpha-D-mannosidases A and B. Biochem J. 1986 Jan 1;233(1):65–72. doi: 10.1042/bj2330065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. D'Agostaro G., Bendiak B., Tropak M. Cloning of cDNA encoding the membrane-bound form of bovine beta 1,4-galactosyltransferase. Eur J Biochem. 1989 Jul 15;183(1):211–217. doi: 10.1111/j.1432-1033.1989.tb14915.x. [DOI] [PubMed] [Google Scholar]
  8. Donaldson J. G., Lippincott-Schwartz J., Bloom G. S., Kreis T. E., Klausner R. D. Dissociation of a 110-kD peripheral membrane protein from the Golgi apparatus is an early event in brefeldin A action. J Cell Biol. 1990 Dec;111(6 Pt 1):2295–2306. doi: 10.1083/jcb.111.6.2295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feizi T. Carbohydrate differentiation antigens: probable ligands for cell adhesion molecules. Trends Biochem Sci. 1991 Mar;16(3):84–86. doi: 10.1016/0968-0004(91)90038-w. [DOI] [PubMed] [Google Scholar]
  10. Feizi T. Demonstration by monoclonal antibodies that carbohydrate structures of glycoproteins and glycolipids are onco-developmental antigens. Nature. 1985 Mar 7;314(6006):53–57. doi: 10.1038/314053a0. [DOI] [PubMed] [Google Scholar]
  11. Fukuda M. N. HEMPAS disease: genetic defect of glycosylation. Glycobiology. 1990 Sep;1(1):9–15. doi: 10.1093/glycob/1.1.9. [DOI] [PubMed] [Google Scholar]
  12. Fukuda M. N., Masri K. A., Dell A., Luzzatto L., Moremen K. W. Incomplete synthesis of N-glycans in congenital dyserythropoietic anemia type II caused by a defect in the gene encoding alpha-mannosidase II. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7443–7447. doi: 10.1073/pnas.87.19.7443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goldberg D. E., Kornfeld S. Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation. J Biol Chem. 1983 Mar 10;258(5):3159–3165. [PubMed] [Google Scholar]
  14. Jackson M. R., Nilsson T., Peterson P. A. Identification of a consensus motif for retention of transmembrane proteins in the endoplasmic reticulum. EMBO J. 1990 Oct;9(10):3153–3162. doi: 10.1002/j.1460-2075.1990.tb07513.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joziasse D. H., Shaper J. H., Van den Eijnden D. H., Van Tunen A. J., Shaper N. L. Bovine alpha 1----3-galactosyltransferase: isolation and characterization of a cDNA clone. Identification of homologous sequences in human genomic DNA. J Biol Chem. 1989 Aug 25;264(24):14290–14297. [PubMed] [Google Scholar]
  16. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  17. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Krayev A. S., Kramerov D. A., Skryabin K. G., Ryskov A. P., Bayev A. A., Georgiev G. P. The nucleotide sequence of the ubiquitous repetitive DNA sequence B1 complementary to the most abundant class of mouse fold-back RNA. Nucleic Acids Res. 1980 Mar 25;8(6):1201–1215. doi: 10.1093/nar/8.6.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Larsen R. D., Rajan V. P., Ruff M. M., Kukowska-Latallo J., Cummings R. D., Lowe J. B. Isolation of a cDNA encoding a murine UDPgalactose:beta-D-galactosyl- 1,4-N-acetyl-D-glucosaminide alpha-1,3-galactosyltransferase: expression cloning by gene transfer. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8227–8231. doi: 10.1073/pnas.86.21.8227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lippincott-Schwartz J., Donaldson J. G., Schweizer A., Berger E. G., Hauri H. P., Yuan L. C., Klausner R. D. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell. 1990 Mar 9;60(5):821–836. doi: 10.1016/0092-8674(90)90096-w. [DOI] [PubMed] [Google Scholar]
  21. Machamer C. E., Mentone S. A., Rose J. K., Farquhar M. G. 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. doi: 10.1073/pnas.87.18.6944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Masibay A. S., Qasba P. K. Expression of bovine beta-1,4-galactosyltransferase cDNA in COS-7 cells. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5733–5737. doi: 10.1073/pnas.86.15.5733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Masri K. A., Appert H. E., Fukuda M. N. 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. doi: 10.1016/s0006-291x(88)80300-0. [DOI] [PubMed] [Google Scholar]
  24. Moremen K. W. Isolation of a rat liver Golgi mannosidase II clone by mixed oligonucleotide-primed amplification of cDNA. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5276–5280. doi: 10.1073/pnas.86.14.5276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moremen K. W., Touster O. Biosynthesis and modification of Golgi mannosidase II in HeLa and 3T3 cells. J Biol Chem. 1985 Jun 10;260(11):6654–6662. [PubMed] [Google Scholar]
  26. Moremen K. W., Touster O., Robbins P. W. Novel purification of the catalytic domain of Golgi alpha-mannosidase II. Characterization and comparison with the intact enzyme. J Biol Chem. 1991 Sep 5;266(25):16876–16885. [PubMed] [Google Scholar]
  27. Moremen K. W., Touster O. Topology of mannosidase II in rat liver Golgi membranes and release of the catalytic domain by selective proteolysis. J Biol Chem. 1986 Aug 15;261(23):10945–10951. [PubMed] [Google Scholar]
  28. Munro S., Pelham H. R. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. doi: 10.1016/0092-8674(87)90086-9. [DOI] [PubMed] [Google Scholar]
  29. Narimatsu H., Sinha S., Brew K., Okayama H., Qasba P. K. Cloning and sequencing of cDNA of bovine N-acetylglucosamine (beta 1-4)galactosyltransferase. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4720–4724. doi: 10.1073/pnas.83.13.4720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nilsson T., Jackson M., Peterson P. A. Short cytoplasmic sequences serve as retention signals for transmembrane proteins in the endoplasmic reticulum. Cell. 1989 Aug 25;58(4):707–718. doi: 10.1016/0092-8674(89)90105-0. [DOI] [PubMed] [Google Scholar]
  31. Nishikawa Y., Pegg W., Paulsen H., Schachter H. Control of glycoprotein synthesis. Purification and characterization of rabbit liver UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I. J Biol Chem. 1988 Jun 15;263(17):8270–8281. [PubMed] [Google Scholar]
  32. Novikoff P. M., Tulsiani D. R., Touster O., Yam A., Novikoff A. B. Immunocytochemical localization of alpha-D-mannosidase II in the Golgi apparatus of rat liver. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4364–4368. doi: 10.1073/pnas.80.14.4364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Paulson J. C., Colley K. J. Glycosyltransferases. Structure, localization, and control of cell type-specific glycosylation. J Biol Chem. 1989 Oct 25;264(30):17615–17618. [PubMed] [Google Scholar]
  34. Paulson J. C. Glycoproteins: what are the sugar chains for? Trends Biochem Sci. 1989 Jul;14(7):272–276. doi: 10.1016/0968-0004(89)90062-5. [DOI] [PubMed] [Google Scholar]
  35. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Päbo S., Bhat B. M., Wold W. S., Peterson P. A. 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. doi: 10.1016/0092-8674(87)90226-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rademacher T. W., Parekh R. B., Dwek R. A. Glycobiology. Annu Rev Biochem. 1988;57:785–838. doi: 10.1146/annurev.bi.57.070188.004033. [DOI] [PubMed] [Google Scholar]
  38. Rees D. J., Ades S. E., Singer S. J., Hynes R. O. Sequence and domain structure of talin. Nature. 1990 Oct 18;347(6294):685–689. doi: 10.1038/347685a0. [DOI] [PubMed] [Google Scholar]
  39. Rothman J. E., Urbani L. J., Brands R. Transport of protein between cytoplasmic membranes of fused cells: correspondence to processes reconstituted in a cell-free system. J Cell Biol. 1984 Jul;99(1 Pt 1):248–259. doi: 10.1083/jcb.99.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Saraste J., Palade G. E., Farquhar M. G. Antibodies to rat pancreas Golgi subfractions: identification of a 58-kD cis-Golgi protein. J Cell Biol. 1987 Nov;105(5):2021–2029. doi: 10.1083/jcb.105.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sarkar M., Hull E., Nishikawa Y., Simpson R. J., Moritz R. L., Dunn R., Schachter H. Molecular cloning and expression of cDNA encoding the enzyme that controls conversion of high-mannose to hybrid and complex N-glycans: UDP-N-acetylglucosamine: alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):234–238. doi: 10.1073/pnas.88.1.234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Shaper N. L., Hollis G. F., Douglas J. G., Kirsch I. R., Shaper J. H. Characterization of the full length cDNA for murine beta-1,4-galactosyltransferase. Novel features at the 5'-end predict two translational start sites at two in-frame AUGs. J Biol Chem. 1988 Jul 25;263(21):10420–10428. [PubMed] [Google Scholar]
  44. Shin J., Dunbrack R. L., Jr, Lee S., Strominger J. L. Signals for retention of transmembrane proteins in the endoplasmic reticulum studied with CD4 truncation mutants. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1918–1922. doi: 10.1073/pnas.88.5.1918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Singer M. F. Highly repeated sequences in mammalian genomes. Int Rev Cytol. 1982;76:67–112. doi: 10.1016/s0074-7696(08)61789-1. [DOI] [PubMed] [Google Scholar]
  46. Tulsiani D. R., Harris T. M., Touster O. Swainsonine inhibits the biosynthesis of complex glycoproteins by inhibition of Golgi mannosidase II. J Biol Chem. 1982 Jul 25;257(14):7936–7939. [PubMed] [Google Scholar]
  47. Tulsiani D. R., Hubbard S. C., Robbins P. W., Touster O. alpha-D-Mannosidases of rat liver Golgi membranes. Mannosidase II is the GlcNAcMAN5-cleaving enzyme in glycoprotein biosynthesis and mannosidases Ia and IB are the enzymes converting Man9 precursors to Man5 intermediates. J Biol Chem. 1982 Apr 10;257(7):3660–3668. [PubMed] [Google Scholar]
  48. Tulsiani D. R., Touster O. Characterization of a novel alpha-D-mannosidase from rat brain microsomes. J Biol Chem. 1985 Oct 25;260(24):13081–13087. [PubMed] [Google Scholar]
  49. Weinstein J., Lee E. U., McEntee K., Lai P. H., Paulson J. C. 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] [Google Scholar]
  50. Wickens M., Stephenson P. Role of the conserved AAUAAA sequence: four AAUAAA point mutants prevent messenger RNA 3' end formation. Science. 1984 Nov 30;226(4678):1045–1051. doi: 10.1126/science.6208611. [DOI] [PubMed] [Google Scholar]
  51. Wickner W. T., Lodish H. F. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. doi: 10.1126/science.4048938. [DOI] [PubMed] [Google Scholar]
  52. Yamamoto F., Marken J., Tsuji T., White T., Clausen H., Hakomori S. Cloning and characterization of DNA complementary to human UDP-GalNAc: Fuc alpha 1----2Gal alpha 1----3GalNAc transferase (histo-blood group A transferase) mRNA. J Biol Chem. 1990 Jan 15;265(2):1146–1151. [PubMed] [Google Scholar]
  53. Yanagisawa K., Resnick D., Abeijon C., Robbins P. W., Hirschberg C. B. A guanosine diphosphatase enriched in Golgi vesicles of Saccharomyces cerevisiae. Purification and characterization. J Biol Chem. 1990 Nov 5;265(31):19351–19355. [PubMed] [Google Scholar]
  54. Yang Y. C., Ciarletta A. B., Temple P. A., Chung M. P., Kovacic S., Witek-Giannotti J. S., Leary A. C., Kriz R., Donahue R. E., Wong G. G. Human IL-3 (multi-CSF): identification by expression cloning of a novel hematopoietic growth factor related to murine IL-3. Cell. 1986 Oct 10;47(1):3–10. doi: 10.1016/0092-8674(86)90360-0. [DOI] [PubMed] [Google Scholar]
  55. Yoshihisa T., Anraku Y. A novel pathway of import of alpha-mannosidase, a marker enzyme of vacuolar membrane, in Saccharomyces cerevisiae. J Biol Chem. 1990 Dec 25;265(36):22418–22425. [PubMed] [Google Scholar]
  56. Yoshihisa T., Anraku Y. Nucleotide sequence of AMS1, the structure gene of vacuolar alpha-mannosidase of Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1989 Sep 15;163(2):908–915. doi: 10.1016/0006-291x(89)92308-5. [DOI] [PubMed] [Google Scholar]

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