The Saccharomyces cerevisiae CWH8 gene is required for full levels of dolichol-linked oligosaccharides in the endoplasmic reticulum and for efficient N-glycosylation

The Saccharomyces cerevisiae CWH8 gene is required for full levels of dolichol-linked... The Saccharomyces cerevisiae mutant cwh8 was previously found to have an anomalous cell wall. Here we show that the cwh8 mutant has an N -glycosylation defect. We found that cwh8 cells were resistant to vanadate and sensitive to hygromycin B, and produced glycoforms of invertase and carboxypeptidase Y with a reduced number of N -chains. We have cloned the CWH8 gene. We found that it was nonessential and encoded a putative transmembrane protein of 239 amino acids. Comparison of the in vitro oligosaccharyl transferase activities of membrane preparations from wild type or cwh8 Δ cells revealed no differences in enzyme kinetic properties indicating that the oligosaccharyl transferase complex of mutant cells was not affected. cwh8 Δ cells also produced normal dolichols and dolichol-linked oligosaccharide intermediates including the full-length form Glc3Man9GlcNAc2. The level of dolichol-linked oligosaccharides in cwh8 Δ cells was, however, reduced to about 20% of the wild type. We propose that inefficient N -glycosylation of secretory proteins in cwh8 Δ cells is caused by an insufficient supply of dolichol-linked oligosaccharide substrate. Key words Key words Calcofluor white hypersensitive carboxypeptidase Y cell wall mutant hygromycin hypersensitive vanadate resistant Abbreviations: Abbreviations CPY carboxypeptidase Y Dol-P dolichyl phosphate Dol-PP dolichyl pyrophosphate Endo H endo-β-N-acetylglucosamidase H ER endoplasmic reticulum OTase oligosaccharyl transferase SGD Saccharomyces Genome Database © 1999 Oxford University Press « Previous | Next Article » Table of Contents This Article Glycobiology (1999) 9 (3): 243-253. » Abstract Free Full Text (HTML) Free Full Text (PDF) Free Classifications Article Services Article metrics Alert me when cited Alert me if corrected Find similar articles Similar articles in Web of Science Similar articles in PubMed Add to my archive Download citation Request Permissions Disclaimer Citing Articles Load citing article information Citing articles via CrossRef Citing articles via Scopus Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by van Berkel, M. A. A. Articles by Klis, F. M. Search for related content PubMed PubMed citation Articles by van Berkel, M. A. A. Articles by Rieger, M. Articles by te Heesen, S. Articles by Ram, A. F. J. Articles by van den Ende, H. Articles by Aebi, M. Articles by Klis, F. M. Related Content Load related web page information Share Email this article CiteULike Delicious Facebook Google+ Mendeley Twitter What's this? Search this journal: Advanced » Current Issue November 2015 25 (11) Alert me to new issues The Journal Submit now! About this journal Rights & Permissions Manuscript submission & review Dispatch date of the next issue This journal is a member of the Committee on Publication Ethics (COPE) We are mobile – find out more Journals Career Network Glycoscience resources Consortium for Functional Glycomics The Official Journal of The Society for Glycobiology Impact factor: 3.147 5-Yr impact factor: 3.212 Editor-in-Chief Robert S. Haltiwanger View full editorial board For Authors Instructions to authors Self-archiving policy Online submission Open access options for authors - visit Oxford Open This journal enables compliance with the NIH Public Access Policy Alerting Services Email table of contents Email Advance Access CiteTrack XML RSS feed Corporate Services Advertising sales Classified Advertising Reprints Supplements var taxonomies = ("SCI01000"); Most Most Read Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds A "Glyconutrient Sham" Formation of the glycan chains in the synthesis of bacterial peptidoglycan Fucose: biosynthesis and biological function in mammals Optimal and consistent protein glycosylation in mammalian cell culture » View all Most Read articles Most Cited Biological roles of oligosaccharides: all of the theories are correct Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites An evolving view of the eukaryotic oligosaccharyltransferase Evolutionary considerations in relating oligosaccharide diversity to biological function Glycosidases of the asparagine-linked oligosaccharide processing pathway » View all Most Cited articles Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department. Online ISSN 1460-2423 - Print ISSN 0959-6658 Copyright © 2015 Oxford University Press Oxford Journals Oxford University Press Site Map Privacy Policy Cookie Policy Legal Notices Frequently Asked Questions Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan Academic & Professional books Children's & Schools Books Dictionaries & Reference Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks International Education Unit Law Medicine Music Online Products & Publishing Oxford Bibliographies Online Oxford Dictionaries Online Oxford English Dictionary Oxford Language Dictionaries Online Oxford Scholarship Online Reference Rights and Permissions Resources for Retailers & Wholesalers Resources for the Healthcare Industry Very Short Introductions World's Classics function fnc_onDomLoaded() { var query_context = getQueryContext(); PF_initOIUnderbar(query_context,":QS:default","","JRN"); PF_insertOIUnderbar(0); }; if (window.addEventListener) { window.addEventListener('load', fnc_onDomLoaded, false); } else if (window.attachEvent) { window.attachEvent('onload', fnc_onDomLoaded); } var gaJsHost = (("https:" == document.location.protocol) ? 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The Saccharomyces cerevisiae CWH8 gene is required for full levels of dolichol-linked oligosaccharides in the endoplasmic reticulum and for efficient N-glycosylation

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Oxford University Press
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Copyright © 2015 Oxford University Press
ISSN
0959-6658
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1460-2423
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glycob;9/3/243
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Abstract

The Saccharomyces cerevisiae mutant cwh8 was previously found to have an anomalous cell wall. Here we show that the cwh8 mutant has an N -glycosylation defect. We found that cwh8 cells were resistant to vanadate and sensitive to hygromycin B, and produced glycoforms of invertase and carboxypeptidase Y with a reduced number of N -chains. We have cloned the CWH8 gene. We found that it was nonessential and encoded a putative transmembrane protein of 239 amino acids. Comparison of the in vitro oligosaccharyl transferase activities of membrane preparations from wild type or cwh8 Δ cells revealed no differences in enzyme kinetic properties indicating that the oligosaccharyl transferase complex of mutant cells was not affected. cwh8 Δ cells also produced normal dolichols and dolichol-linked oligosaccharide intermediates including the full-length form Glc3Man9GlcNAc2. The level of dolichol-linked oligosaccharides in cwh8 Δ cells was, however, reduced to about 20% of the wild type. We propose that inefficient N -glycosylation of secretory proteins in cwh8 Δ cells is caused by an insufficient supply of dolichol-linked oligosaccharide substrate. Key words Key words Calcofluor white hypersensitive carboxypeptidase Y cell wall mutant hygromycin hypersensitive vanadate resistant Abbreviations: Abbreviations CPY carboxypeptidase Y Dol-P dolichyl phosphate Dol-PP dolichyl pyrophosphate Endo H endo-β-N-acetylglucosamidase H ER endoplasmic reticulum OTase oligosaccharyl transferase SGD Saccharomyces Genome Database © 1999 Oxford University Press « Previous | Next Article » Table of Contents This Article Glycobiology (1999) 9 (3): 243-253. » Abstract Free Full Text (HTML) Free Full Text (PDF) Free Classifications Article Services Article metrics Alert me when cited Alert me if corrected Find similar articles Similar articles in Web of Science Similar articles in PubMed Add to my archive Download citation Request Permissions Disclaimer Citing Articles Load citing article information Citing articles via CrossRef Citing articles via Scopus Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by van Berkel, M. A. A. Articles by Klis, F. M. Search for related content PubMed PubMed citation Articles by van Berkel, M. A. A. Articles by Rieger, M. Articles by te Heesen, S. Articles by Ram, A. F. J. Articles by van den Ende, H. Articles by Aebi, M. Articles by Klis, F. M. Related Content Load related web page information Share Email this article CiteULike Delicious Facebook Google+ Mendeley Twitter What's this? Search this journal: Advanced » Current Issue November 2015 25 (11) Alert me to new issues The Journal Submit now! About this journal Rights & Permissions Manuscript submission & review Dispatch date of the next issue This journal is a member of the Committee on Publication Ethics (COPE) We are mobile – find out more Journals Career Network Glycoscience resources Consortium for Functional Glycomics The Official Journal of The Society for Glycobiology Impact factor: 3.147 5-Yr impact factor: 3.212 Editor-in-Chief Robert S. Haltiwanger View full editorial board For Authors Instructions to authors Self-archiving policy Online submission Open access options for authors - visit Oxford Open This journal enables compliance with the NIH Public Access Policy Alerting Services Email table of contents Email Advance Access CiteTrack XML RSS feed Corporate Services Advertising sales Classified Advertising Reprints Supplements var taxonomies = ("SCI01000"); Most Most Read Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds A "Glyconutrient Sham" Formation of the glycan chains in the synthesis of bacterial peptidoglycan Fucose: biosynthesis and biological function in mammals Optimal and consistent protein glycosylation in mammalian cell culture » View all Most Read articles Most Cited Biological roles of oligosaccharides: all of the theories are correct Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites An evolving view of the eukaryotic oligosaccharyltransferase Evolutionary considerations in relating oligosaccharide diversity to biological function Glycosidases of the asparagine-linked oligosaccharide processing pathway » View all Most Cited articles Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department. Online ISSN 1460-2423 - Print ISSN 0959-6658 Copyright © 2015 Oxford University Press Oxford Journals Oxford University Press Site Map Privacy Policy Cookie Policy Legal Notices Frequently Asked Questions Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan Academic & Professional books Children's & Schools Books Dictionaries & Reference Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks International Education Unit Law Medicine Music Online Products & Publishing Oxford Bibliographies Online Oxford Dictionaries Online Oxford English Dictionary Oxford Language Dictionaries Online Oxford Scholarship Online Reference Rights and Permissions Resources for Retailers & Wholesalers Resources for the Healthcare Industry Very Short Introductions World's Classics function fnc_onDomLoaded() { var query_context = getQueryContext(); PF_initOIUnderbar(query_context,":QS:default","","JRN"); PF_insertOIUnderbar(0); }; if (window.addEventListener) { window.addEventListener('load', fnc_onDomLoaded, false); } else if (window.attachEvent) { window.attachEvent('onload', fnc_onDomLoaded); } var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); try { var pageTracker = _gat._getTracker("UA-189672-16"); pageTracker._setDomainName(".oxfordjournals.org"); pageTracker._trackPageview(); } catch(err) {}

Journal

GlycobiologyOxford University Press

Published: Mar 1, 1999

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