Document Detail


A mutation causing a reduced level of expression of six beta4-galactosyltransferase genes is the basis of the Lec19 CHO glycosylation mutant.
MedLine Citation:
PMID:  14567696     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
JaeHoon Lee; Sung-Hae Park; Subha Sundaram; T Shantha Raju; Nancy L Shaper; Pamela Stanley
Related Documents :
1908946 - Decreased clastogenicity of dinitropyrenes in chinese hamster lung (chl) subclone cells...
10770596 - Isolation of liver oval cells from hamsters treated with diethylnitrosamine and 2-acety...
7739616 - Toxicity of camptothecin to chinese hamster cells containing 5-hydroxymethyl-2'-deoxyur...
8231826 - Persistence of sister chromatid exchange by 9-beta-d-arabinofuranosyladenine in chinese...
25166766 - Cell block technique as an additional tool in the diagnosis of ameloblastoma.
22369946 - Identification of mir-508-3p and mir-509-3p that are associated with cell invasion and ...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochemistry     Volume:  42     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2003-10-21     Completed Date:  2003-12-02     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  12349-57     Citation Subset:  IM    
Affiliation:
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
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:
R01 36434//PHS HHS
Chemical
Reg. No./Substance:
0/DNA Primers; 0/Lectins; EC 2.4.1.-/Galactosyltransferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Kinetics of fibrinopeptide release by thrombin as a function of CaCl2 concentration: different susce...
Next Document:  Channel-opening kinetics of GluR2Q(flip) AMPA receptor: a laser-pulse photolysis study.