S2.12 Transcriptional regulation of murine?1,4-galactosyltransferase

Harduin-Lepers, Anne; Shaper, Joël H.; Shaper, Nancy L.

doi:10.1007/bf01209855

Cited by 18 publications

(45 citation statements)

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“…Tissue-specific alternative splicing has been reported for several glycosyltransferases, including ␣-2,3-(9) and ␣-2,6-sialyltransferases (30, 31), ␤-1,4-Gal-transferase (7,8), and GlcNActransferases I (32) and V (33). The 5Ј-flanking regions of the ␣-2,3-sialyltransferase isoforms contain heterogeneous transcriptional start sites and different transcriptional regulation sequences (9).…”

Section: Resultsmentioning

confidence: 99%

Tissue-specific Regulation of Mouse Core 2 ॆ-1,6-N-Acetylglucosaminyltransferase

Sekine

Nara

Suzuki

1997

Journal of Biological Chemistry

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Mouse kidney ␤-1,6-GlcNAc-transferase (GNT) is the key enzyme for the synthesis of a glycosphingolipid (Gal␤1-4(Fuc␣1-3)GlcNAc␤1-6(Gal␤1-3)GalNAc␤1-3Gal␣1-4Gal␤1-4Glc␤1-ceramide) that contains the Le X trisaccharide epitope at its nonreducing terminus. The expression of this glycolipid in the kidney is polymorphic; it is expressed in BALB/c but not DBA/2 mice; and a single autosomal gene (Gsl5) is responsible for this polymorphism. We report here the cDNA sequence that encodes the kidney GNT of BALB/c mice, which possess a wild-type Gsl5 gene. The deduced amino acid sequence exhibits 84% identity to that of human core 2 ␤-1,6-GlcNAc-transferase, which suggests that kidney GNT is a mouse homologue of human core 2 ␤-1,6-GlcNAc-transferase. The GNT mRNA is expressed abundantly in the kidney, but was not detected in other BALB/c organs or in the kidneys of DBA/2 mice by Northern blot analysis. In addition, we were able to clone and sequence another homologous cDNA from the submandibular gland. The two sequences differ only in their 5-untranslated region. The submandibular gland type of cDNA was detected in various organs of DBA/2 mice by reverse transcription-polymerase chain reaction, which indicates that the submandibular gland type is ubiquitous and that its expression is not regulated by the Gsl5 gene. Results obtained using the long accurate polymerase chain reaction method indicate that the GNT gene is ϳ45 kilobases long, and the order of the exons from the 5-end is exon 1 of the kidney type, exon 1 of the ubiquitous type, exon 2, and exon 3. Exons 2 and 3 are present in both transcripts, and the translated region is in exon 3. These data suggest that the expression of GNT is regulated by an alternative splicing mechanism and also probably by tissue-specific enhancers and that Gsl5 regulates the expression of GNT only in the kidney.Carbohydrate chains of cell-surface glycoconjugates play important roles in cell-cell and cell-matrix communication (1). The diversity of the carbohydrate structures provides a basis for cell-specific recognition. The expression of carbohydrate chains is highly regulated and changes during embryogenesis, differentiation, and oncogenic transformation (2). The regulatory process may be mediated by many different gene products, including glycosyltransferases (3, 4), transcription factors (5-10), nucleotide sugar transporters (11-14), kinases and phosphatases that act on transferases (15,16), and other genes. The objective of our studies is to understand the genetic basis and mechanisms that regulate the expression of carbohydrates.We have identified an autosomal mouse gene (Gsl5) that controls the expression of GlcNAc␤1-6(Gal␤1-3)GalNAc␤1-3Gb 3 Cer and its elongated glycolipids by regulating ␤-1,6-GlcNAc-transferase (GNT) 1 activity in the kidney (17). DBA/2 mice are not able to express detectable levels of GNT activity or the glycolipids containing GlcNAc␤1-6(Gal␤1-3)GalNAc␤1-3Gb 3 Cer as a core structure because of a defect of the Gsl5 gene. To elucidate the role of Gsl5, the mouse k...

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Section: Resultsmentioning

confidence: 99%

Tissue-specific Regulation of Mouse Core 2 ॆ-1,6-N-Acetylglucosaminyltransferase

Sekine

Nara

Suzuki

1997

Journal of Biological Chemistry

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“…For example, ␤4GalT-1 is up-regulated in lactating mammary glands (16), whereas ␤4GalT-2 is not. 2 Furthermore, mice deficient in ␤4GalT-1 do not produce lactose in milk (17,18).…”

Section: ␤4-galactosyltransferase (␤4galt)mentioning

confidence: 99%

“…ϩ RNA, 0.5 g of oligo(dT) [12][13][14][15][16][17][18] primer, and 0.05 g of random hexamer were heated to 70°C for 10 min and slowly cooled to room temperature before adding 200 -400 units of Superscript II reverse transcriptase together with first strand buffer (Life Technologies, Inc.), 0.5 mM dATP, 0.5 mM dCTP, 0.5 mM dGTP, 0.5 mM dTTP, 10 mM dithiothreitol, and 1 unit/l RNasin (Promega). Reactions were incubated for 50 min at 42°C, heated at 70°C for 15 min, and stored at Ϫ20°C.…”

Section: Reverse Transcription (Rt)-pcr-for Reverse Transcription 2 mentioning

confidence: 99%

Chinese Hamster Ovary (CHO) Cells May Express Six β4-Galactosyltransferases (β4GalTs)

Lee

Sundaram

Shaper

et al. 2001

Journal of Biological Chemistry

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Six ␤4-galactosyltransferase (␤4GalT) genes have been cloned from mammalian sources. We show that all six genes are expressed in the Gat ؊ 2 line of Chinese hamster ovary cells (Gat ؊ 2 CHO). Two independent mutants termed Pro ؊ 5Lec20 and Gat ؊ 2Lec20, previously selected for lectin resistance, were found to have a galactosylation defect. Radiolabeled biantennary Nglycans synthesized by Pro ؊ 5Lec20 were proportionately less ricin-bound than similar species from parental CHO cells, and Lec20 cell extracts had a markedly reduced ability to transfer Gal to GlcNAc-terminating acceptors. Northern blot analysis revealed a severe reduction in ␤4GalT-1 transcripts in Pro ؊ 5Lec20 cells. The Gat ؊ 2Lec20 mutant expressed ␤4GalT-1 transcripts of reduced size due to a 311-base pair deletion in the ␤4GalT-1 gene coding region. Northern analysis with probes from the remaining five ␤4GalT genes revealed that Gat ؊ 2 CHO and Gat ؊ 2Lec20 cells express all six ␤4GalT genes. Unexpectedly, the ␤4GalT-6 gene is not expressed in either Pro ؊ 5 or Pro ؊ 5Lec20 cells. Thus, in addition to a deficiency in ␤4GalT-1, Pro ؊ 5Lec20 cells lack ␤4GalT-6. Nevertheless, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry data of N-glycans released from cellular glycoproteins showed that both the ␤4GalT-1 ؊ (Gat ؊ 2Lec20) and ␤4GalT-1 ؊ /␤4GalT-6 ؊ (Pro ؊ 5Lec20) mutants have a similar Gal deficiency, affecting neutral and sialylated bi-, tri-, and tetraantennary N-glycans. By contrast, glycolipid synthesis was normal in both mutants. Therefore, ␤4GalT-1 is a key enzyme in the galactosylation of Nglycans, but is not involved in glycolipid synthesis in CHO cells. ␤4GalT-6 contributes only slightly to the galactosylation of N-glycans and is also not involved in CHO cell glycolipid synthesis. These CHO glycosylation mutants provide insight into the variety of in vivo substrates of different ␤4GalTs. They may be used in glycosylation engineering and in investigating roles for ␤4GalT-1 and ␤4GalT-6 in generating specific glycan ligands.

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“…In mouse somatic tissues, the start site for the 4.1-kb transcript is used predominantly, and the expression from this start site is controlled by a promoter containing multiple Sp1-binding sites (74,75). The only exception to this pattern is found in the mammary gland during lactation, where there is a switch to the preferential use of the start site for the 3.9-kb transcript, and expression from this start site is controlled by the promoter regulated by lactating mammary gland-restricted transcription factors (74,75). The most distal transcription start site is used exclusively during the late stages of spermatogenesis (76,77).…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Regulation of the Human β-1,4-Galactosyltransferase V Gene in Cancer Cells

Satô

Furukawa

2004

Journal of Biological Chemistry

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S2.12 Transcriptional regulation of murine?1,4-galactosyltransferase

Cited by 18 publications

References 0 publications

Tissue-specific Regulation of Mouse Core 2 ॆ-1,6-N-Acetylglucosaminyltransferase

Tissue-specific Regulation of Mouse Core 2 ॆ-1,6-N-Acetylglucosaminyltransferase

Chinese Hamster Ovary (CHO) Cells May Express Six β4-Galactosyltransferases (β4GalTs)

Transcriptional Regulation of the Human β-1,4-Galactosyltransferase V Gene in Cancer Cells

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