Benzyl-N-acetyl-α-d-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation

Delannoy, Philippe; Kim, Isabelle; Emery, Nancy C.; Bolós, Carme de; Verbert, André; Degand, Pierre; Huet, Guillemette

doi:10.1007/bf00702335

Cited by 57 publications

(55 citation statements)

References 28 publications

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“…However, unmyristoylated annexin XIIIb was not bound to the membrane but exclusively present in the cytosolic fraction (50). Beyond the requirement of the myristoyl moiety for mem-brane binding (50), our data suggest that in HT-29 5M12 cells, the association of membrane-bound annexin XIIIb with lipid microdomains involves its interaction with sialylated epitopes, as we showed that GalNAc␣-O-bn inhibits sialylation of glycoproteins and likely glycolipids by the two sialyltransferases ST3Gal I and ST3Gal IV (22,23).…”

Section: Discussionmentioning

confidence: 57%

“…In this cell line, GalNAc␣-O-bn was found to inhibit primarily the terminal sialylation by the sialyltransferases ST3Gal I and ST3Gal IV, and we suggested that sialylation of apical glycoproteins may act as a targeting signal for apical delivery (19,22,23). The goal of this work was to examine the anterograde traffic in cloned HT-29 5M12 cells material of GalNAc␣-O-bn-treated cells shifted in fraction 2 of lower buoyant density.…”

Section: Discussionmentioning

confidence: 97%

“…In a previous work, we showed that GalNAc␣-O-bn was highly converted into the benzyldisaccharide Gal␤1-3GalNAc␣-O-bn, which acts as a potent competitive inhibitor of ␣2,3-sialyltransferase ST3Gal I and involved in the terminal elongation of O-linked glycans (22). Thereafter, we reported that GalNAc␣-O-bn was extensively metabolized beyond Gal␤1-3GalNAc␣-O-bn, showing that the glycosylation of endogenous substrates by several glycosyltransferases could be inhibited, and in particular the sialylation of N-glycans by ␣2,3-sialyltransferase ST3Gal IV (21,23).…”

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confidence: 99%

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1-Benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside Blocks the Apical Biosynthetic Pathway in Polarized HT-29 Cells

Delacour

Gouyer

Leteurtre

et al. 2003

Journal of Biological Chemistry

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In previous work we reported that long term treatment of polarized HT-29 cells by 1-benzyl-2-acetamido-2-deoxy-␣-D-galactopyranoside (GalNAc␣-O-bn) induced undersialylation and intracellular distribution of apical glycoproteins such as dipeptidyl peptidase IV (DPP-IV), and we suggested therefore that sialylation could act as an apical targeting signal. In this work, the apical direct biosynthetic route was studied after transfection of polarized enterocyte-like HT-29 5M12 cloned cells with a murine cDNA coding for a soluble form of DPP-IV, which was secreted into the apical medium. A 24-h treatment of transfected cells by GalNAc␣-O-bn markedly inhibited the apical secretion and the sialylation of this soluble murine DPP-IV, which became blocked inside the cell. A similar short GalNAc␣-O-bn treatment also induced an intracellular distribution of both endogenous transmembrane DPP-IV and proteins involved in the regulation of the apical trafficking such as the apical t-SNARE syntaxin-3 and the raft-associated protein annexin XIIIb, whereas the basolateral t-SNARE syntaxin-4 kept its normal localization. These apical membrane proteins moved efficiently from trans-Golgi network to apical carrier vesicles but failed to be transported from carrier vesicles to the apical plasma membrane. Isolation of membrane microdomains showed that GalNAc␣-O-bn induced the formation of abnormal lipid-rich microdomains in comparison to normal rafts, as shown by their lower buoyant density and their depletion in annexin XIIIb. In conclusion, GalNAc␣-O-bn blocks the anterograde traffic to the apical surface of polarized HT-29 cells at the transport level or docking/fusion level of carrier vesicles.

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Section: Discussionmentioning

confidence: 57%

Section: Discussionmentioning

confidence: 97%

mentioning

confidence: 99%

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1-Benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside Blocks the Apical Biosynthetic Pathway in Polarized HT-29 Cells

Delacour

Gouyer

Leteurtre

et al. 2003

Journal of Biological Chemistry

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“…This effect was shown to be cell-specific and connected to the inhibition of the sialylation of mucins by ST3Gal I, a glycosyltransferase highly expressed in HT-29 cells. 50,52 Here, we observed a specific inhibition in the localization of MUC1 and MUC5AC at the surface of GalNAc␣-O-bn treated HT-29 5M21 cells cultured on collagen, whereas the localization of E-cadherin was not affected. This observation showed that the gain of Ecadherin function in GalNAc␣-O-bn treated HT-29 5M21 cells could not be simply attributed to an inhibition of the glycosylation of mucins but to a more complex mechanism resulting in the loss of expression of MUC1 and MUC5AC at the periphery of the cell.…”

Section: Immunofluorescence Analysis Of the Distribution Of E-cadherimentioning

confidence: 66%

Requirement of both mucins and proteoglycans in cell‐cell dissociation and invasiveness of colon carcinoma HT‐29 cells

Truant

Bruyneel

Gouyer

et al. 2003

Intl Journal of Cancer

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Key words: colon carcinoma cells; E-cadherin; mucins; heparan sulfate proteoglycansEpithelial E-cadherin, a component of the adherens junctions, is crucial for the organization and maintenance of differentiated epithelia. 1 The extracellular NH2-terminal domain of this transmembrane glycoprotein mediates cell-cell adhesion in a homophilic and Ca 2ϩ -dependent way. The COOH-terminal domain binds to the actin cytoskeleton via ␣-catenin and ␤-catenin or plakoglobin. 2 Studies in cancer cell lines have shown that changes in the structure or in the expression of the components of the E-cadherin/␤-catenin complex result in the suppression of cell-cell adhesion and in the expression of an invasive phenotype. [3][4][5][6][7][8] In cancer, the maintenance of the epithelia is lost and dissociation of cells is associated with metastatic dissemination. This phenomenon can occur through a decrease in the expression level of E-cadherin and, regarding colon cancer, such decrease has been related to the tumor grade and clinical outcome. 9 At the genomic level, loss of E-cadherin expression does not occur as a result of mutation of E-cadherin gene, whereas mutations of ␤-catenin were found in a small proportion of colon cancers. 10 -12 In another way, E-cadherin-mediated cell-cell adhesion may be modulated by the mucin-like glycoprotein MUC1, which is often overexpressed in cancer. MUC1, also termed episialin, is a membrane-associated glycoprotein composed of a membrane anchor and a mucin-like ectodomain. 13 In human ZR-75-1 breast cancer cells, the cytoplasmic domain of MUC1 was shown to bind ␤-catenin, leading to a decrease of E-cadherin-catenin complex and an anti-adhesion effect. 14 On the other hand, MUC1 was shown to cause steric hindrance of adhesion molecules like E-cadherin, through its large negatively charged extracellular domain, which contains many oligosaccharide side chains. 15,16 Proteoglycans were also shown to disturb the function of E-cadherin through steric hindrance in variants of the murine mammary gland cell line NMuMG and the canine epithelial kidney cell line MDCK. 17 During colorectal carcinogenesis, the expression of MUC1 was found upregulated, particularly in later stages of the disease. 18 -20 On the other hand, de novo appearance of a secreted mucin normally found in the stomach, MUC5AC, has been described in colon carcinomas. [21][22][23] Thus, mucinous differentiated cells are present in colorectal carcinomas and more particularly in mucinous carcinomas and in signet ring cell carcinomas. Until now, the significance of a differentiation characteristic into a mucin-secreting phenotype with regard to the tumor development is still unknown.To investigate the role of mucins in the invasive properties of colon carcinoma cells, the HT-29 cell line is an appropriate model, as the cells of this cell line express different phenotypes: besides a majority of undifferentiated cells (Ͼ95%), a small proportion of cells presents a capacity to differentiate into a mucinous phenotype or into an enterocyte-like ...

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“…Previous studies have suggested that O-glycans may influence protein stability, as is thought to be the case in the human disease, familial tumoral calcinosis (26,(31)(32). Other studies in cell culture using chemical inhibitors of O-glycan extension have demonstrated that alterations in O-glycan formation can affect secretion and trafficking of certain proteins (33)(34)(35)(36). However, these inhibitors also affect other glycosylation pathways, making specific conclusions about the role of mucin-type O-glycans difficult.…”

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confidence: 98%

An O-Glycosyltransferase Promotes Cell Adhesion during Development by Influencing Secretion of an Extracellular Matrix Integrin Ligand

Zhang

Tran

Hagen

2010

Journal of Biological Chemistry

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Protein secretion and localization are crucial during eukaryotic development, establishing local cell environments as well as mediating cell interactions, signaling, and adhesion. In this study, we demonstrate that the glycosyltransferase, pgant3, specifically modulates integrin-mediated cell adhesion by influencing the secretion and localization of the integrin ligand, Tiggrin. We demonstrate that Tiggrin is normally O-glycosylated and localized to the basal matrix where the dorsal and ventral cell layers adhere in wild type Drosophila wings. In pgant3 mutants, Tiggrin is no longer O-glycosylated and fails to be properly secreted to this basal cell layer interface, resulting in disruption of integrin-mediated cell adhesion in the wing. pgant3-mediated effects are dependent on enzymatic activity, as mutations that form a stable protein yet abrogate O-glycosyltransferase activity result in Tiggrin accumulation within the dorsal and ventral cells comprising the wing. Our results provide the first in vivo evidence for the role of O-glycosylation in the secretion of specific extracellular matrix proteins, thus altering the composition of the cellular "microenvironment" and thereby modulating developmentally regulated cell adhesion events. As alterations in cell adhesion are a hallmark of cancer progression, this work provides insight into the long-standing association between aberrant O-glycosylation and tumorigenesis.

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Benzyl-N-acetyl-α-d-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation

Cited by 57 publications

References 28 publications

1-Benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside Blocks the Apical Biosynthetic Pathway in Polarized HT-29 Cells

1-Benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside Blocks the Apical Biosynthetic Pathway in Polarized HT-29 Cells

Requirement of both mucins and proteoglycans in cell‐cell dissociation and invasiveness of colon carcinoma HT‐29 cells

An O-Glycosyltransferase Promotes Cell Adhesion during Development by Influencing Secretion of an Extracellular Matrix Integrin Ligand

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