| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 274, Issue 15, 10439-10450, April 9, 1999
From the Department of Cell Biology, Albert Einstein College of
Medicine, New York, New York, 10461 and ¶ Cytel Corporation,
La Jolla, California 92121
The LEC11 Chinese hamster ovary (CHO)
gain-of-function mutant expresses an 
(1,3)fucosyltransferase
((1,3)Fuc-T) activity that generates the LeX,
sialyl-LeX, and VIM-2 glycan determinants and has been
extensively used for studies of E-selectin ligand specificity. In order
to identify regulatory mechanisms that control (1,3)Fuc-T expression
in mammals, mechanisms of FUT gene expression were
investigated in LEC11 cells and two new, independent mutants, LEC11A
and LEC11B. Northern and ribonuclease protection analyses, using probes
that span the coding region of a cloned CHO FUT gene,
detected transcripts in each LEC11 mutant but not in CHO cells or other
gain-of-function CHO mutants that express a different (1,3)Fuc-T
activity. Coding region sequence analysis and (1,3)Fuc-T acceptor
specificity comparisons with recombinant human Fuc-TV and Fuc-TVI
showed that the cloned FUT gene is orthologous to the human
FUT6 gene. Southern analyses identified two closely related
FUT6 genes in the Chinese hamster, whose evolutionary
relationships are discussed. The blots showed that rearrangements had
occurred in LEC11A and LEC11 genomic DNA, consistent with a
cis mechanism of FUT6 gene activation in these
mutants. By contrast, somatic cell hybrid analyses revealed that LEC11B
cells express FUT6 gene transcripts due to the loss of a
trans-acting, negative regulatory factor. Sequencing of
reverse transcriptase-polymerase chain reaction products identified
unique 5'- and 3'-untranslated region sequences in FUT6
gene transcripts from each LEC11 mutant. Northern and Southern analyses
with gene-specific probes showed that LEC11A cells express only the
cgFUT6A gene (where cg is Cricetulus griseus),
whereas LEC11 and LEC11B cells express only the cgFUT6B
gene. In LEC11A × LEC11B hybrid cells, the cgFUT6A
gene was predominantly expressed, as predicted if a
trans-acting negative regulatory factor functions to
suppress cgFUT6B gene expression in CHO cells. This factor
is predicted to be a cell type-specific regulator of FUT6
gene expression in mammals.
This article has been cited by other articles:
|
|
 |
|
  O. W. Liu, M. J. S. Kelly, E. D. Chow, and H. D. Madhani Parallel {beta}-Helix Proteins Required for Accurate Capsule Polysaccharide Synthesis and Virulence in the Yeast Cryptococcus neoformans Eukaryot. Cell, April 1, 2007; 6(4): 630 - 640. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Chen, J. Tang, and P. Stanley Suppressors of {alpha}(1,3)fucosylation identified by expression cloning in the LEC11B gain-of-function CHO mutant Glycobiology, March 1, 2005; 15(3): 259 - 269. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 S. K. Patnaik, B. Potvin, and P. Stanley LEC12 and LEC29 Gain-of-Function Chinese Hamster Ovary Mutants Reveal Mechanisms for Regulating VIM-2 Antigen Synthesis and E-selectin Binding J. Biol. Chem., November 26, 2004; 279(48): 49716 - 49726. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. J. Thomas, K. Panneerselvam, D. T. Beattie, M. D. Picard, B. Xu, C. W. Rittershaus, H. C. Marsh Jr., R. A. Hammond, J. Qian, T. Stevenson, et al. Production of a complement inhibitor possessing sialyl Lewis X moieties by in vitro glycosylation technology Glycobiology, October 1, 2004; 14(10): 883 - 893. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Hong and P. Stanley Lec3 Chinese Hamster Ovary Mutants Lack UDP-N-acetylglucosamine 2-Epimerase Activity Because of Mutations in the Epimerase Domain of the Gne Gene J. Biol. Chem., December 26, 2003; 278(52): 53045 - 53054. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Nishihara, H. Iwasaki, K. Nakajima, A. Togayachi, Y. Ikehara, T. Kudo, Y. Kushi, A. Furuya, K. Shitara, and H. Narimatsu {alpha}1,3-Fucosyltransferase IX (Fut9) determines Lewis X expression in brain Glycobiology, June 1, 2003; 13(6): 445 - 455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Sherwood, D. A. Upchurch, M. R. Stroud, W. C. Davis, and E. H. Holmes A highly conserved His-His motif present in {alpha}1->3/4fucosyltransferases is required for optimal activity and functions in acceptor binding Glycobiology, October 1, 2002; 12(10): 599 - 606. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Dupuy, A. Germot, M. Marenda, R. Oriol, A. Blancher, R. Julien, and A. Maftah {alpha}1,4-Fucosyltransferase Activity: A Significant Function in the Primate Lineage has Appeared Twice Independently Mol. Biol. Evol., June 1, 2002; 19(6): 815 - 824. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Norgard-Sumnicht, X. Bai, J. D. Esko, A. Varki, and A. E. Manzi Exploring the outcome of genetic modifications of glycosylation in cultured cell lines by concurrent isolation of the major classes of vertebrate glycans Glycobiology, July 1, 2000; 10(7): 691 - 700. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Cao, Z. Zhao, R. Mohan, J. Alroy, P. Stanley, and N. Panjwani Role of the Lewisx Glycan Determinant in Corneal Epithelial Cell Adhesion and Differentiation J. Biol. Chem., June 8, 2001; 276(24): 21714 - 21723. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
