| A mutation causing a reduced level of expression of six beta4-galactosyltransferase genes is the basis of the Lec19 CHO glycosylation mutant. | |
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MedLine Citation:
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PMID: 14567696 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To identify factors required for the synthesis of complex glycans, we have isolated Chinese hamster ovary (CHO) cell mutants resistant to plant lectins. We previously identified Lec19 CHO cells as resistant to the Gal-binding lectins ricin, abrin, and modeccin and hypersensitive to the toxicity of other lectins that bind Gal, including L-PHA and E-PHA. Here we show that Lec19 cell extracts have a decreased ability to transfer Gal to simple sugar, oligosaccharide, and glycopeptide acceptors, particularly to biantennary, GlcNAc-terminated acceptors. Ricin(II)-agarose lectin affinity chromatography, oligomapping, and monosaccharide analyses provided evidence that Lec19 N-glycans have fewer Gal residues than CHO N-glycans. MALDI-TOF mass spectra of N-glycans released from Lec19 cell glycoproteins by peptide N-glycanase F revealed species with the predicted masses of neutral N-glycans with few Gal residues. Such truncated species are essentially absent from CHO cell glycoproteins. However, the complement of fully galactosylated or sialylated bi-, tri-, and tetra-antennary N-glycans was largely equivalent in Lec19 and CHO cells. In addition, the coding region sequences of the beta4GalT-1, -T-2, -T-3, -T-4, -T-5, and -T-6 genes were identical in CHO and Lec19 cells. However, Northern analyses revealed an approximately 2-4-fold reduction in the level of transcripts of all six beta4GalT genes in Lec19 cells. Since the recessive Lec19 phenotype is the result of a loss-of-function mutation, the combined data predict the existence of a trans-acting regulator of the steady-state level of transcripts that derive from these six mammalian beta4GalT genes. |
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Authors:
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JaeHoon Lee; Sung-Hae Park; Subha Sundaram; T Shantha Raju; Nancy L Shaper; Pamela Stanley |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biochemistry Volume: 42 ISSN: 0006-2960 ISO Abbreviation: Biochemistry Publication Date: 2003 Oct |
Date Detail:
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Created Date: 2003-10-21 Completed Date: 2003-12-02 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0370623 Medline TA: Biochemistry Country: United States |
Other Details:
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Languages: eng Pagination: 12349-57 Citation Subset: IM |
Affiliation:
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Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Blotting, Northern CHO Cells Chromatography, High Pressure Liquid Chromatography, Ion Exchange Cricetinae DNA Primers Galactosyltransferases / genetics*, metabolism Glycosylation Lectins / metabolism Mutation* Protein Binding Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| Grant Support | |
ID/Acronym/Agency:
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R01 36434//PHS HHS |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/Lectins; EC 2.4.1.-/Galactosyltransferases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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