Skip Navigation


Glycobiology Advance Access originally published online on October 30, 2002
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
13/1/43    most recent
cwg003v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (30)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Chen, W.
Right arrow Articles by Stanley, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, W.
Right arrow Articles by Stanley, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Glycobiology, 2003, Vol. 13, No. 1 43-50
© 2003 Oxford University Press

Five Lec1 CHO cell mutants have distinct Mgat1 gene mutations that encode truncated N-acetylglucosaminyltransferase I

Wei Chen and Pamela Stanley1

Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave., New York, NY 10461, USA

Received on August 2, 2002; revised on August 21, 2002; accepted on August 23, 2002

Lec1 CHO cell mutants lack N-acetylglucosaminyltransferase I (GlcNAc-TI) activity and do not synthesize complex or hybrid N-glycans. The origins of six independent lec1 mutations are shown to reside in the coding region of the Mgat1 gene, proving that GlcNAc-TI is mutated in Lec1 mutants. One mutant has Mgat1 gene transcripts of reduced size, whereas the others possess transcripts of approximately normal size and amount containing a unique insertion or transition mutation that leads to a premature stop codon in the Mgat1 gene coding region. The lec1 mutation in the Lec3.2.8.1 mutant, a line used to generate minimally glycosylated membrane glycoproteins for X-ray crystallography, is a G insertion that leads to a nonsense codon after amino acid 391. The Pro-Lec1.3C line from the ATCC and in laboratory stocks, a line used widely for diverse purposes, possesses a C insertion in the Mgat1 gene coding exon, causing a frame shift and producing a stable, truncated ~24-kDa product. Mgat1 gene mutations were confirmed by sequencing genomic DNA PCR products. Mutant cDNAs were reverted by site-directed mutagenesis and shown to confer wild-type lectin binding and GlcNAc-TI activity on Lec1 transfectants. Surprisingly, three Mgat1 gene nucleotide changes previously reported in Pro-Lec1.3C cells (Puthalakath et al. [1996] J. Biol. Chem., 271, 27818–27822) were not detected in this study. These Lec1 mutants provide a novel cohort for investigating the effects on Golgi trafficking and kin recognition of deletion mutants of GlcNAc-TI expressed at endogenous rather than nonphysiological levels.

1 To whom correspondence should be addressed; e-mail: stanley{at}aecom.yu.edu


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 GlycobiologyHome page
J. T Aguilan, S. Sundaram, E. Nieves, and P. Stanley
Mutational and functional analysis of Large in a novel CHO glycosylation mutant
Glycobiology, September 1, 2009; 19(9): 971 - 986.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Crispin, V. T. Chang, D. J. Harvey, R. A. Dwek, E. J. Evans, D. I. Stuart, E. Y. Jones, J. M. Lord, R. A. Spooner, and S. J. Davis
A Human Embryonic Kidney 293T Cell Line Mutated at the Golgi {alpha}-Mannosidase II Locus
J. Biol. Chem., August 7, 2009; 284(32): 21684 - 21695.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. D. Nightingale, M. E. F. Frayne, S. Clasper, S. Banerji, and D. G. Jackson
A Mechanism of Sialylation Functionally Silences the Hyaluronan Receptor LYVE-1 in Lymphatic Endothelium
J. Biol. Chem., February 6, 2009; 284(6): 3935 - 3945.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
W. Zhou and H.-M. Tsai
N-Glycans of ADAMTS13 modulate its secretion and von Willebrand factor cleaving activity
Blood, January 22, 2009; 113(4): 929 - 935.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Stahl, K. Uemura, C. Ge, S. Shi, Y. Tashima, and P. Stanley
Roles of Pofut1 and O-Fucose in Mammalian Notch Signaling
J. Biol. Chem., May 16, 2008; 283(20): 13638 - 13651.
[Abstract] [Full Text] [PDF]

Home page
 JEMHome page
T. Nochi, Y. Yuki, A. Matsumura, M. Mejima, K. Terahara, D.-Y. Kim, S. Fukuyama, K. Iwatsuki-Horimoto, Y. Kawaoka, T. Kohda, et al.
A novel M cell specific carbohydrate-targeted mucosal vaccine effectively induces antigen-specific immune responses
J. Exp. Med., November 26, 2007; 204(12): 2789 - 2796.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
H. Tateno, N. Uchiyama, A. Kuno, A. Togayachi, T. Sato, H. Narimatsu, and J. Hirabayashi
A novel strategy for mammalian cell surface glycome profiling using lectin microarray
Glycobiology, October 1, 2007; 17(10): 1138 - 1146.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Shi, C. Ge, Y. Luo, X. Hou, R. S. Haltiwanger, and P. Stanley
The Threonine That Carries Fucose, but Not Fucose, Is Required for Cripto to Facilitate Nodal Signaling
J. Biol. Chem., July 13, 2007; 282(28): 20133 - 20141.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
V. I. Otto, E. Damoc, L. N. Cueni, T. Schurpf, R. Frei, S. Ali, N. Callewaert, A. Moise, J. A. Leary, G. Folkers, et al.
N-Glycan structures and N-glycosylation sites of mouse soluble intercellular adhesion molecule-1 revealed by MALDI-TOF and FTICR mass spectrometry
Glycobiology, November 1, 2006; 16(11): 1033 - 1044.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
M. Crispin, D. J. Harvey, V. T. Chang, C. Yu, A. R. Aricescu, E. Y. Jones, S. J. Davis, R. A. Dwek, and P. M. Rudd
Inhibition of hybrid- and complex-type glycosylation reveals the presence of the GlcNAc transferase I-independent fucosylation pathway
Glycobiology, August 1, 2006; 16(8): 748 - 756.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Sarkar, P. A. Leventis, C. I. Silvescu, V. N. Reinhold, H. Schachter, and G. L. Boulianne
Null Mutations in Drosophila N-Acetylglucosaminyltransferase I Produce Defects in Locomotion and a Reduced Life Span
J. Biol. Chem., May 5, 2006; 281(18): 12776 - 12785.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
S. K. Patnaik, B. Potvin, S. Carlsson, D. Sturm, H. Leffler, and P. Stanley
Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells
Glycobiology, April 1, 2006; 16(4): 305 - 317.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
S. Subramanian, R. Parthasarathy, S. Sen, E. T. Boder, and D. E. Discher
Species- and cell type-specific interactions between CD47 and human SIRP{alpha}
Blood, March 15, 2006; 107(6): 2548 - 2556.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
S. Wopereis, E. Morava, S. Grunewald, M. Adamowicz, K. M. L. C. Huijben, D. J. Lefeber, and R. A. Wevers
Patients with unsolved congenital disorders of glycosylation type II can be subdivided in six distinct biochemical groups
Glycobiology, December 1, 2005; 15(12): 1312 - 1319.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
R. R. Novoa, G. Calderita, P. Cabezas, R. M. Elliott, and C. Risco
Key Golgi Factors for Structural and Functional Maturation of Bunyamwera Virus
J. Virol., September 1, 2005; 79(17): 10852 - 10863.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. K. Patnaik and P. Stanley
Mouse Large Can Modify Complex N- and Mucin O-Glycans on {alpha}-Dystroglycan to Induce Laminin Binding
J. Biol. Chem., May 27, 2005; 280(21): 20851 - 20859.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
P. Stanley, S. Sundaram, J. Tang, and S. Shi
Molecular analysis of three gain-of-function CHO mutants that add the bisecting GlcNAc to N-glycans
Glycobiology, January 1, 2005; 15(1): 43 - 53.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
S. Shi, S. A. Williams, A. Seppo, H. Kurniawan, W. Chen, Z. Ye, J. D. Marth, and P. Stanley
Inactivation of the Mgat1 Gene in Oocytes Impairs Oogenesis, but Embryos Lacking Complex and Hybrid N-Glycans Develop and Implant
Mol. Cell. Biol., November 15, 2004; 24(22): 9920 - 9929.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
V. I. Otto, T. Schurpf, G. Folkers, and R. D. Cummings
Sialylated Complex-type N-Glycans Enhance the Signaling Activity of Soluble Intercellular Adhesion Molecule-1 in Mouse Astrocytes
J. Biol. Chem., August 20, 2004; 279(34): 35201 - 35209.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. H. Merry, R. J. C. Gilbert, D. A. Shore, L. Royle, O. Miroshnychenko, M. Vuong, M. R. Wormald, D. J. Harvey, R. A. Dwek, B. J. Classon, et al.
O-Glycan Sialylation and the Structure of the Stalk-like Region of the T Cell Co-receptor CD8
J. Biol. Chem., July 11, 2003; 278(29): 27119 - 27128.
[Abstract] [Full Text] [PDF]


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.