Functional Conservation of Subfamilies of Putative UDP-N-acetylgalactosamine:Polypeptide N-Acetylgalactosaminyltransferases inDrosophila, Caenorhabditis elegans, and Mammals

Schwientek, Tilo; Bennett, Eric; Flores, Carlos; Thacker, John; Hollmann, Martin; Reis, Celso A.; Behrens, Jane; Mandel, Ulla; Keck, Birgit; Schäfer, Mireille; Haselmann, Kim F.; Zubarev, Roman A.; Roepstorff, Peter; Burchell, Joy; Taylor‐Papadimitriou, Joyce; Hollingsworth, Michael A.; Clausen, Henrik

doi:10.1074/jbc.m202684200

Cited by 172 publications

(152 citation statements)

References 46 publications

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“…Human pp-GalNAc-T3 and -T6 is highly or restrictively expressed in pancreas and testis, and in placenta and trachea, respectively (6). pp-GalNAc-T11 is restrictively expressed in kidney (9). pp-GalNAc-T9 was found to be expressed mostly in brain (8).…”

Section: Discussionmentioning

confidence: 99%

Cloning and Characterization of a New Human UDP-N-Acetyl-α-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase, Designated pp-GalNAc-T13, That Is Specifically Expressed in Neurons and Synthesizes GalNAc α-Serine/Threonine Antigen

Zhang

Iwasaki

Wang

et al. 2003

Journal of Biological Chemistry

127

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To date, 10 members of the UDP-N-acetyl-␣-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase (pp-GalNAc-T) family have been cloned and analyzed in human. In this study, we cloned and analyzed a novel human pp-GalNAc-T from an NT2 cell cDNA library, and we named it pp-GalNAc-T13. In amino acid sequences, pp-GalNAc-T13 was highly homologous, showing 84.3% identity, to pp-GalNAc-T1. Real time PCR analysis revealed pp-GalNAc-T13 to be highly and restrictively expressed in the brain and present at very low or undetectable levels in other tissues, in contrast to the ubiquitous expression of pp-GalNAc-T1. pp-GalNAc-T13 was abundantly expressed in all neuroblastoma cells examined and primary cultured neurons but not in glioblastoma cells and primary cultured astrocytes. pp-GalNAc-T13 exhibited much stronger activity to transfer GalNAc to mucin peptides, such as Muc5Ac and MUC7, than did pp-GalNAc-T1. In addition, pp-GalNAc-T13 differed in substrate specificity to pp-GalNAc-T1. pp-GalNAc-T13 was able to form a triplet Tn epitope, three consecutive GalNAc-Ser/Thr structures, on peptides encoded in syndecan-3, a proteoglycan expressed in neurons. pp-GalNAc-T13-deficient mice have been established in a previous work. Immunohistochemical study showed a remarkable decrease in Tn antigen expression in the cerebellum of the pp-GalNAc-T13 knockout mouse. pp-GalNAc-T13 would be a major enzyme responsible for the synthesis of O-glycan and specifically the Tn antigen epitope in neurons.O-Linked glycosylation of mucin is initiated by the transfer of N-acetylgalactosamine with an ␣-linkage to a serine or threonine residue in protein. The GalNAc residue attached to the peptide is usually extended to form more complex O-glycan structures by the action of multiple glycosyltransferases. The addition of GalNAc is controlled by multiple members of the pp-GalNAc-T 1 (EC 2.4.1.41) family. To date, 10 distinct members have been identified in human (pp-GalNAc-T1, -T2, -T3, -T4, -T6 to -T9, -T11, and -T12) (1-10) and 7 in rodent (ppGaN- Tase-T1, -T2, -T3, -T4, -T5, -T7, -T10 2 ) (11-17). All share a highly homologous primary sequence, 40 -60% homology at the amino acid level, to each other particularly in the predicted catalytic domain. Each member exhibits different substrate specificity toward peptide sequences. Thus, the positions of O-glycan in proteins are determined by the substrate specificity of each pp-GalNAc-T. The genes encoding these enzymes are distributed at different genomic localizations on chromosomes and have distinct genomic structures (4).pp-GalNAc-T1, which was the first pp-GalNAc-T to be cloned from bovine tissue (1), is the best characterized of the members. It shows activity toward Muc1, Muc2, Muc5Ac, and Muc7 peptides (4, 18). Peptide sequences derived from these mucins were used to determine the site of the O-glycosylation of the Muc2 and Muc5Ac mucin tandem repeat region by pp-GalNAc-T1 (19 -22).Many pp-GalNAc-Ts, including pp-GalNAc-T1, were found to * This work was performed as a part of the R&D Project o...

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Section: Discussionmentioning

confidence: 99%

Cloning and Characterization of a New Human UDP-N-Acetyl-α-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase, Designated pp-GalNAc-T13, That Is Specifically Expressed in Neurons and Synthesizes GalNAc α-Serine/Threonine Antigen

Zhang

Iwasaki

Wang

et al. 2003

Journal of Biological Chemistry

127

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“…Some flies with mutations related to proteoglycan, dally (34), sugarless, toutvelu, and sulfateless (2), have demonstrated defects in signaling of the growth factors including Wingless, Decapentaplegic, Hedgehog, and fibroblast growth factors. Recently, one recessive lethal mutant has been reported to have a missense mutation causing a reduction of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase activity (35,36). Considering the recent progress made in studies of Drosophila glycans as mentioned above, D. melanogaster will become a powerful tool for analysis of the biological roles of glycans.…”

Section: Identification Of the Drosophila Proteoglycan ␤14-mentioning

confidence: 99%

Proteoglycan UDP-Galactose:β-Xylose β1,4-Galactosyltransferase I Is Essential for Viability inDrosophila melanogaster

Takemae

Ueda

Okubo

et al. 2003

Journal of Biological Chemistry

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Heparan and chondroitin sulfates play essential roles in growth factor signaling during development and share a common linkage tetrasaccharide structure, GlcA␤1,3Gal␤1,3Gal␤1,4Xyl␤1-O-Ser. In the present study, we identified the Drosophila proteoglycan UDP-galactose:␤-xylose ␤1,4-galactosyltransferase I (d␤4GalTI), and determined its substrate specificity. The enzyme transferred a Gal to the -␤-xylose (Xyl) residue, confirming it to be the Drosophila ortholog of human proteoglycan UDP-galactose:␤-xylose ␤1,4-galactosyltransferase I. Then we established UAS-d␤4GalTI-IR fly lines containing an inverted repeat of d␤4GalTI ligated to the upstream activating sequence (UAS) promoter, a target of GAL4, and observed the F 1 generation of the cross between the UAS-d␤4GalTI-IR fly and the Act5C-GAL4 fly. In the F 1 , double-stranded RNA of d␤4GalTI is expressed ubiquitously under the control of a cytoplasmic actin promoter to induce the silencing of the d␤4GalTI gene. The expression of the target gene was disrupted specifically, and the degree of interference was correlated with phenotype. The lethality among the progeny proved that ␤4GalTI is essential for viability. This study is the first to use reverse genetics, RNA interference, to study the Drosophila glycosyltransferase systematically.

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“…However, it also remains possible that, for some functions, core 1 O-linked glycans are functionally redundant with other Olinked or N-linked glycans. Although Drosophila also contain several ppGalNAcTs, expressed in overlapping patterns (Ten Hagen et al, 2003b;Tian and Ten Hagen, 2006), mutation of at least one ppGalNAcT, l(2)35a, is lethal (Schwientek et al, 2002;Ten Hagen and Tran, 2002), and mutant animals have tracheal defects (Tian and Ten Hagen, 2007b).…”

Section: Introductionmentioning

confidence: 99%

Requirement for a core 1 galactosyltransferase in the Drosophila nervous system

Lin

Reddy

Irvine

2008

Developmental Dynamics

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Mucin type O-glycosylation is a widespread modification of eukaryotic proteins, but its functional requirements remain incompletely understood. It is initiated by the attachment of N-acetylgalactosamine (GalNAc) to Ser or Thr residues, and then elongated by additional sugars. We have examined requirements for mucin-type glycosylation in Drosophila by characterizing the expression and phenotypes of core 1 galactosyltransferases (core 1 GalTs), which elongate O-GalNAc by adding galactose in a ␤1,3 linkage. Drosophila encode several putative core 1 GalTs, each expressed in distinct patterns. CG9520 (C1GalTA) is expressed in the amnioserosa and central nervous system. A null mutation in C1GalTA is lethal, and mutant animals exhibit a striking morphogenetic defect in which the ventral nerve cord is greatly elongated and the brain hemispheres are misshapen. Lectin staining and blotting experiments confirmed that C1GalTA contributes to the synthesis of Gal-␤1,3-GalNAc in vivo. Our results identify a role for mucin-type Oglycosylation during neural development in Drosophila. Developmental Dynamics 237:3703-3714, 2008.

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Functional Conservation of Subfamilies of Putative UDP-N-acetylgalactosamine:Polypeptide N-Acetylgalactosaminyltransferases inDrosophila, Caenorhabditis elegans, and Mammals

Cited by 172 publications

References 46 publications

Cloning and Characterization of a New Human UDP-N-Acetyl-α-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase, Designated pp-GalNAc-T13, That Is Specifically Expressed in Neurons and Synthesizes GalNAc α-Serine/Threonine Antigen

Cloning and Characterization of a New Human UDP-N-Acetyl-α-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase, Designated pp-GalNAc-T13, That Is Specifically Expressed in Neurons and Synthesizes GalNAc α-Serine/Threonine Antigen

Proteoglycan UDP-Galactose:β-Xylose β1,4-Galactosyltransferase I Is Essential for Viability inDrosophila melanogaster

Requirement for a core 1 galactosyltransferase in the Drosophila nervous system

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