Abstract
All asparagine-linked glycans (N-glycans) on the eukaryotic glycoproteins are primarily derived from dolichol-linked oligosaccharides (DLO), synthesized on the rough endoplasmic reticulum membrane. We have previously reported cloning and identification of the human gene, HMT-1, which encodes chitobiosyldiphosphodolichol beta-mannosyltransferase (β1,4-MT) involved in the early assembly of DLO. Considering that N-glycosylation is one of the most ubiquitous post-translational modifications for many eukaryotic proteins, the HMT-1 could be postulated as one of the housekeeping genes, but its transcriptional regulation remains to be investigated. Here we screened a 1 kb region upstream from HMT-1 open reading frame (ORF) for transcriptionally regulatory sequences by using chloramphenicol acetyl transferase (CAT) assay, and found that the region from −33 to −1 positions might act in HMT-1 transcription at basal level and that the region from −200 to −42 should regulate its transcription either positively or negatively. In addition, results with CAT assays suggested the possibility that two GATA-1 motifs and an Sp1 motif within a 200 bp region upstream from HMT-1 ORF might significantly upregulate HMT-1 transcription. On the contrary, the observations obtained from site-directed mutational analyses revealed that an NF-1/AP-2 overlapping motif located at −148 to −134 positions should serve as a strong silencer. The control of the HMT-1 transcription by these motifs resided within the 200 bp region could partially explain the variation of expression level among various human tissues, suggesting availability and importance of this region for regulatory role in HMT-1 expression.
Similar content being viewed by others
Abbreviations
- DLO:
-
Dolichol-linked oligosaccharide
- MT:
-
Mannosyltransferase
- rER:
-
Rough endoplasmic reticulum
- β1,4-MT:
-
Beta 1,4-mannosyltransferase
- HMT-1:
-
Human mannosyltransferase I
- kb:
-
Kilo base pair
- ORF:
-
Open reading frame
- CAT:
-
Chloramphenicol acetyl transferase
- GATA-1:
-
GATA-binding factor 1
- Sp1:
-
Specificity protein 1
- bp:
-
Base pair
- NF-1:
-
Nuclear factor 1
- AP-2:
-
Activating protein 2
- Man:
-
Mannose
- Dol:
-
Dolichol
- GlcNAc:
-
N-Acetylglucosamine
- UDP:
-
Uridine diphosphate
- GDP:
-
Guanosine diphosphate
- Glc:
-
Glucose
- Dol-P:
-
Dolichyl phosphate
- DPAGT1:
-
Dolichyl-phosphate (UDP-N-acetylglucosamine) N-acetylglucosaminephosphotransferase 1
- GlcNAc-1-P:
-
N-Acetylglucosamine-1-phosphate
- ALG1:
-
Asparagine-linked glycosylation 1
- CDG-I:
-
Congenital disorder of glycosylation type I
- α1,3/α1,6-MT:
-
Alpha 1,3/alpha 1,6-mannosyltransferase
- α1,2-MT:
-
Alpha 1,2-mannosyltransferase
- DMEM:
-
Dulbecco’s modified Eagle medium
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- G3PDH:
-
Glyceraldehyde 3-phosphate dehydrogenase
- DEPC:
-
Diethylpyrocarbonate
- PBS:
-
Phosphate buffered saline
- EDTA:
-
Ethylenediaminetetraacetic acid
- OD:
-
Optical density
- ONPG:
-
Ortho-nitrophenyl-beta-d-galactopyranoside
- TLC:
-
Thin layer chromatography
- SE:
-
Standard error
- USF:
-
Upstream stimulatory factor
- AP-1:
-
Activating protein 1
- AP-4:
-
Activating protein 4
- Dex:
-
Dexamethasone
- RSV:
-
Rous sarcoma virus
- MMTV:
-
Mouse mammary tumor virus
- LTR:
-
Long terminal repeat
- Ets-1:
-
E-twenty-six-1
- NF-kB:
-
Nuclear factor-kappa B
- STAT5a:
-
Signal transducer and activator of transcription 5a
- TCF-1:
-
Transcription factor 1
- Alg7p:
-
Asparagine-linked glycosylation 7 protein
- Alg14p:
-
Asparagine-linked glycosylation 14 protein
References
Adamowicz M, Chmielinska E, Kaluzny L, Bittner G, Sarnowska-Wroczynska I, Timal S, Morava E, Lehle L, Wevers RA, Lefeber DJ, Sykut-Cegielska J (2011) Clinical and biochemical characterization of the second CDGIJ (DPAGT1-CDG) patient. J Inherit Metab Dis 34:S181
Aebi M (2013) N-linked protein glycosylation in the ER. Biochim Biophys Acta 1833:2430–2437
Albright CF, Robbins PW (1990) The sequence and transcript heterogeneity of the yeast gene ALG1, an essential mannosyltransferase involved in N-glycosylation. J Biol Chem 265:7042–7049
Basiri K, Belaya K, Liu WW, Maxwell S, Sedghi M, Beeson D (2013) Clinical features in a large Iranian family with a limb-girdle congenital myasthenic syndrome due to a mutation in DPAGT1. Neuromuscul Disord 23:469–472
Buczkowska A, Swiezewska A, Lefeber DJ (2015) Genetic defect in dolichol metabolism. J Inherit Metab Dis 38:157–169
Cantagrel V, Lefeber DJ (2011) From glycosylation disorders to dolichol biosynthesis defects: a new class of metabolic diseases. J Inherit Metab Dis 34:859–867
Carrera IA, Matthijs G, Perez B, Cerdá CP (2012) DPAGT1-CDG: report of a patient with fetal hypokinesia phenotype. Am J Med Genet A 158A:2027–2030
Couto JR, Huffaker TC, Robbins PW (1984) Cloning and expression in Escherichia coli of a yeast mannosyltransferase from the asparagine-linked glycosylation pathway. J Biol Chem 259:378–382
Cui Y, Narayanan CS, Zhou J, Kumar A (1998) Exon-I is involved in positive as well as negative regulation of human angiotensinogen gene expression. Gene 224:97–107
de Koning TJ, Toet M, Dorland L, de Vries LS, van den Berg IET, Duran M, Poll-The BT (1998) Recurrent nonimmune hydrops fetal is associated with carbohydrate-deficient glycoprotein syndrome. J Inherit Metab Dis 21:681–682
Dupré T, Vuillaumier-Barrot S, Chantret I, Yayé HS, Le Bizec C, Afenjar A, Altuzarra C, Barnérias C, Burglen L, de Lonlay P, Feillet F, Napuri S, Seta N, Moore SEH (2010) Guanosine diphosphate- mannose:GlcNAc2-PP-dolichol mannosyltransferase deficiency (congenital disorders of glycosylation type Ik): five new patients and seven novel mutations. J Med Genet 47:729–735
Eckert D, Buhl S, Weber S, Jäger R, Schorle H (2005) The AP-2 family of transcription factors. Genome Biol 6:246.1–246.8
Elbein AD (1984) Inhibitors of the biosynthesis and processing of N-linked oligosaccharides. CRC Crit Rev Biochem 16:21–49
Finlay-Schultz J, Canastar A, Short M, El Gazzar M, Coughlan C, Leonard S (2011) Transcriptional repression of the α7 nicotinic acetylcholine receptor subunit gene (CHRNA7) by activating protein-2α(AP-2α). J Biol Chem 286:42123–42132
Fischer KD, Haese A, Nowock J (1993) Cooperation of GATA-1 and Sp1 can result in synergistic transcriptional activation or interference. J Biol Chem 268:23915–23923
Furusawa M, Taira T, Iguchi-Ariga SMM, Ariga H (2003) Molecular cloning of the mouse AMY-1 gene and identification of the synergistic activation of the AMY-1 promoter by GATA-1 and Sp1. Genomics 81:221–233
Gao X-D, Nishikawa A, Dean N (2004) Physical interactions between the Alg1, Alg2, and Alg11 mannosyltransferases of the endoplasmic reticulum. Glycobiology 14:559–570
Gronostajski RM (2000) Roles of the NFI/CTF gene family in transcription and development. Gene 249:31–45
Grubenmann CE, Frank CG, Hülsmeier AJ, Schollen E, Matthijs G, Mayatepek E, Berger EG, Aebi M, Hennet T (2004) Deficiency of the first mannosylation step in the N-glycosylation pathway causes congenital disorder of glycosylation type Ik. Hum Mol Genet 13:535–542
Haeuptle MA, Hennet T (2009) Congenital disorders of glycosylation: an update on defects affecting the biosynthesis of dolichol-linked oligosaccharides. Hum Mutat 30:1628–1641
Helander A, Stödberg T, Jaeken J, Matthijs G, Eriksson M, Eggertsen G (2013) Dolichol kinase deficiency (DOLK-CDG) with a purely neurological presentation caused by a novel mutation. Mol Genet Metab 110:342–344
Helenius A, Aebi M (2001) Intracellular functions of N-linked glycans. Science 291:2364–2369
Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of Escherichia coli with plasmids. Gene 96:23–28
Jaeken J (2010) Congenital disorders of glycosylation. Ann NY Acad Sci 1214:190–198
Jung P, Menssen A, Mayr D, Hermeking H (2008) AP4 encodes a c-MYC-inducible repressor of p21. Proc Natl Acad Sci USA 105:15046–15051
Kean EL, Wei Z, Anderson VE, Zhang N, Sayre LM (1999) Regulation of the biosynthesis of N-acetylglucosaminylpyrophosphoryldolichol, feedback and product inhibition. J Biol Chem 274:34072–34082
Kim MY, Jeong BC, Lee JH, Kee HJ, Kook H, Kim NS, Kim YH, Kim JK, Ahn KY, Kim KK (2006) A repressor complex, AP4 transcription factor and geminin, negatively regulates expression of target genes in nonneuronal cells. Proc Natl Acad Sci USA 103:13074–13079
Kranz C, Denecke J, Lehle L, Sohlbach K, Jeske S, Meinhardt F, Rossi R, Gudowius S, Marquardt T (2004) Congenital disorder of glycosylation type Ik (CDG-Ik): a defect of mannosyltransferase I. Am Hum Genet 74:545–551
Kranz C, Jungeblut C, Denecke J, Erlekotte A, Sohlback C, Debus V, Kehl HG, Harms E, Reith A, Reichel S, Grobe H, Hammersen G, Schwarzer U, Marquardt T (2007) A defect in dolichol phosphate biosynthesis causes a new inherited disorder with death in early infancy. Am J Hum Genet 80:433–440
Ku WC, Chiu SK, Chen YJ, Huang HH, Wu WG, Chen YJ (2009) Complementary quantitative proteomics reveals that transcription factor AP-4 mediates E-box-dependent complex formation for transcriptional repression of HDM2. Mol Cell Proteomics 8:2034–2050
Kukuruzinska MA, Bergh MLE, Jackson BJ (1987) Protein glycosylation in yeast. Ann Rev Biochem 56:915–944
Lennon K, Pretel R, Kesselheim R, te Heesen S, Kukuruzinska MA (1995) Proliferation-dependent differential regulation of dolichol pathway genes in Saccharomyces cerevisiae. Glycobiology 5:633–642
Lieu MT, Ng BG, Rush JS, Wood T, Basehore MJ, Hegde M, Chang RC, Abdenur JE, Freeze HH, Wang RY (2013) Severe, fatal multisystem manifestations in a patient with dolichol kinase-congenital disorder of glycosylation. Mol Genet Metab 110:484–489
Liu Y, Bernard HU, Apt D (1997) NFI-B3, a Novel transcriptional repressor of the nuclear factor I family, is generated by alternative RNA processing. J Biol Chem 272:10739–10745
Lu J, Takahashi T, Ohoka A, Nakajima K, Hashimoto R, Miura N, Tachikawa H, Gao X-D (2012) Alg14 organizes the formation of a multi-glycosyltransferase complex involved in initiation of lipid-linked oligosaccharide biosynthesis. Glycobiology 22:504–516
Ma SL, Tang NL, Tam CW, Lui VW, Lam LC, Chiu HF, Driver JA, Pastorino L, Lu KP (2012) A PIN1 polymorphism that prevents its suppression by AP4 associates with delayed onset of Alzheimer’s disease. Neurobiol Aging 33:804–813
Marek KW, Vijay IK, Marth JD (1999) A recessive deletion in the GlcNAc-1-phosphotransferase gene results in peri-implantation embryonic lethality. Glycobiology 9:1263–1271
Merika M, Orkin SH (1995) Functional synergy and physical interactions of the erythroid transcription factor GATA-1 with the Kruppel family proteins Sp1 and EKLF. Mol Cell Biol 15:2437–2447
Mitchell DL, DiMario JX (2010) AP-2α suppresses skeletal myoblast proliferation and represses fibroblast growth factor receptor 1 promoter activity. Exp Cell Res 316:194–202
Morava E, Vodopiutz J, Lefeber DJ, Janecke AR, Schmidt WM, Lechner S, Item CB, Sykut- Cegielska J, Adamowicz M, Wierzba J, Zhang ZH, Mihalek I, Stockler S, Bodamer OA, Lehle L, Wevers RA (2012) Defining the phenotype in congenital disorder of glycosylation due to ALG1 mutations. Pediatrics 130:e1034–e1039
Noffz C, Keppler-Ross S, Dean N (2009) Hetero-oligomeric interactions between early glycosyltransferases of the dolichol cycle. Glycobiology 19:472–478
Regal L, van Hasselt PM, Foulquier F, Cuppen I, Prinsen HCMT, Jansen K, Keldermans L, De Meirleir L, Matthijs G, Jaeken J (2015) ALG11-CDG: three novel mutations and further characterization of the phenotype. Mol Genet Metab Rep 2:16–19
Rind N, Schmeiser V, Thiel C, Absmanner B, Lübbehusen J, Hocks J, Apeshiotis N, Wilichowski E, Lehle L, Körner C (2010) A severe human metabolic disease caused by deficiency of the endoplasmic mannosyltransferase hALG11 leads to congenital disorder of glycosylation-Ip. Hum Mol Genet 19:1413–1424
Rohlfing AK, Rust S, Reunert J, Tirre M, Du Chesne I, Wemhoff S, Meinhardt F, Hartmann H, Das AM, Marquardt T (2014) ALG1-CDG: a new case with early fatal outcome. Gene 534:345–351
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, NY
Schwarz F, Aebi M (2011) Mechanisms and principles of N-linked protein glycosylation. Curr Opin Struct Biol 21:576–582
Schwarz M, Thiel C, Lübbehusen J, Dorland B, de Koning T, von Figura K, Lehle L, Körner C (2004) Deficiency of GDP-Man:GlcNAc2-PP-dolichol mannosyltransferase causes congenital disorder of glycosylation type Ik. Am J Hum Genet 74:472–481
Sengupta PK, Bouchie MP, Kukuruzinska MA (2010) N-Glycosylation gene DPAGT1 is a target of the Wnt/β-catenin signaling pathway. J Biol Chem 285:31164–31173
Sengupta PK, Bouchie MP, Nita-Lazar M, Yang HY, Kukuruzinska MA (2013) Coordinate regulation of N-glycosylation gene DPAGT1, canonical Wnt signaling and E-cadherin adhesion. J Cell Sci 126:484–496
Snow TM, Woods CW, Woods AG (2012) Congenital disorder of glycosylation: a case presentation. Adv Neonatal Care 12:96–100
Su AI, Wiltshire T, Batalov S, Lapp H, Ching KA, Block D, Zhang J, Soden R, Hayakawa M, Kreiman G, Cooke MP, Walker JR, Hogenesch JB (2004) A gene atlas of the mouse and human protein-encoding transcriptomes. Proc Natl Acad Sci USA 101:6062–6067
Takahashi T, Gao X-D (2012) Physical interactions among human glycosyltransferases involved in dolichol-linked oligosaccharide biosynthesis. Trends Glycosci Glycotechnol 24:65–77
Takahashi T, Honda R, Nishikawa Y (2000) Cloning of the human cDNA which can complement the defect of the yeast mannosyltransferase I-deficient mutant alg1. Glycobiology 10:321–327
Thiel C, Schwarz M, Peng J, Grzmil M, Hasilik M, Braulke T, Kohlschütter A, von Figura K, Lehle L, Körner C (2003) A new type of congenital disorders of glycosylation (CDG-Ii) provides new insights into the early steps of dolichol-linked oligosaccharide biosynthesis. J Biol Chem 278:22498–22505
Thiel C, Rind N, Popovici D, Hoffmann GF, Hanson K, Conway RL, Adamski CR, Butler E, Scanion R, Lambert M, Apeshiotis N, Thiel C, Matthijs G, Körner C (2012) Improved diagnostics lead to identification of three new patients with congenital disorder of glycosylation-Ip. Hum Mutat 33:485–487
Timal S, Hoischen A, Lehle L, Adamowicz M, Huijben K, Sykut-Cegielska J, Paprocka J, Jamroz E, van Spronsen FJ, Körner C, Gilissen C, Rodenburg RJ, Eidhof I, Van den Heuvel L, Thiel C, Wevers RA, Morava E, Veltman J, Lefeber DJ (2012) Gene identification in the congenital disorders of glycosylation type I by whole-exome sequencing. Hum Mol Genet 21:4151–4161
Varelas X, Bouchie MP, Kukuruzinska MA (2014) Protein N-glycosylation in oral cancer: dysregulated cellular networks among DPAGT1, E-cadherin adhesion and canonical Wnt signaling. Glycobiology 24:579–591
Varki A (1993) Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3:97–130
Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11
Wu X, Rush JS, Karaoglu D, Krasnewich D, Lubinsky MS, Waechter CJ, Gilmore R, Freeze HH (2003) Deficiency of UDP-GlcNAc:dolichol phosphate N-acetylglucosamine-1 phosphate transferase (DPAGT1) causes a novel congenital disorder of glycosylation type Ij. Hum Mutat 22:144–150
Wu C, Orozco C, Boyer J, Leglise M, Goodale J, Batalov S, Hodge CL, Haase J, Janes J, Huss JW, Su AI (2009) BioGPS: an extensible and customizable portal for querying and organizing gene annotation resources. Genome Biol 10:R130.1–R130.8
Würde AE, Reunert J, Rust S, Hertzberg C, Haverkämper S, Nürnberg G, Nürnberg P, Lehle L, Rossi R, Marquardt T (2012) Congenital disorder of glycosylation type Ij (CDG-Ij, DPAGT1-CDG): extending the clinical and molecular spectrum of a rare disease. Mol Genet Metab 105:634–641
Zhang XL, Qu XJ, Vijay IK (2003) STAT5a regulates the GlcNAc-1-phosphate transferase gene transcription and expression. Cell Physiol Biochem 13:85–92
Acknowledgments
We thank Prof. Y. Nishikawa (Tokai University) for kindly advising us. This study was supported in part by the Grants-in-Aid for Scientific Research on Priority Area No. 12760066 from the Ministry of Education, Science, Sports and Culture of Japan, and the research-funds from School of Engineering at Tokai University in 2000.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Takahashi, T., Nedachi, T., Etoh, T. et al. Identification and characterization of transcriptional control region of the human beta 1,4-mannosyltransferase gene. Cytotechnology 69, 417–434 (2017). https://doi.org/10.1007/s10616-015-9929-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-015-9929-y