HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Nguyen, T. T. Articles by Shur, B. D. Search for Related Content PubMed PubMed Citation Articles by Nguyen, T. T. Articles by Shur, B. D. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 269, Issue 45, 28000-28009, 11, 1994

Expressing murine beta 1,4-galactosyltransferase in HeLa cells produces a cell surface galactosyltransferase-dependent phenotype

TT Nguyen, DA Hinton and BD Shur
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.

Beta 1,4-Galactosyltransferase is traditionally viewed as a biosynthetic component of the Golgi complex, but a portion of galactosyltransferase is also expressed on the cell surface, where it has been suggested to function as a receptor for extracellular oligosaccharide ligands. Although results from a variety of studies are consistent with a cell adhesion function for galactosyltransferase, the most rigorous test of surface galactosyltransferase function is to produce a surface galactosyltransferase-dependent phenotype in cells that normally express negligible levels of surface galactosyltransferase. In agreement with previous reports, human HeLa cells were found to express low levels of galactosyltransferase on their surface and, therefore, were stably transfected with cDNAs encoding murine galactosyltransferase. Murine galactosyltransferase was expressed both within the presumed Golgi complex and on the cell surface, as assayed by enzyme activity and with antiserum raised against the bacterially expressed murine enzyme. HeLa cell transfectants adhered more strongly to their extracellular substrates than did control transfectants, as evidenced by a flatter morphology in culture and a more rapid spreading upon plating. In contrast, cell spreading was low and similar among all cell types when plated on extracellular substrates that did not contain binding sites for galactosyltransferase. Antibodies and Fab fragments against recombinant murine galactosyltransferase inhibited the increased cell spreading characteristic of galactosyltransferase transfectants, as did soluble recombinant galactosyltransferase and a variety of galactosyltransferase perturbants. Thus, expression of heterologous galactosyltransferase produces a surface galactosyltransferase- dependent phenotype, confirming its function as a cell adhesion molecule.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				H. J. Hathaway, S. C. Evans, D. H. Dubois, C. I. Foote, B. H. Elder, and B. D. Shur Mutational analysis of the cytoplasmic domain of {beta}1,4-galactosyltransferase I: influence of phosphorylation on cell surface expression J. Cell Sci., November 1, 2003; 116(21): 4319 - 4330. [Abstract] [Full Text] [PDF]

				M. J. Wassler, C. I. Foote, I. H. Gelman, and B. D. Shur Functional interaction between the SSeCKS scaffolding protein and the cytoplasmic domain of {beta}1,4-galactosyltransferase J. Cell Sci., March 8, 2002; 114(12): 2291 - 2300. [Abstract] [Full Text] [PDF]

				E. G. Berger Ectopic localizations of Golgi glycosyltransferases Glycobiology, February 1, 2002; 12(2): 29R - 36R. [Abstract] [Full Text] [PDF]

				M. Wassler and B. Shur Clustering of cell surface (beta)1,4-galactosyltransferase I induces transient tyrosine phosphorylation of focal adhesion kinase and loss of stress fibers J. Cell Sci., January 1, 2000; 113(2): 237 - 245. [Abstract] [PDF]

				Q Lu and B. Shur Sperm from beta 1,4-galactosyltransferase-null mice are refractory to ZP3-induced acrosome reactions and penetrate the zona pellucida poorly Development, January 10, 1997; 124(20): 4121 - 4131. [Abstract] [PDF]

				H. Hathaway and B. Shur Mammary gland morphogenesis is inhibited in transgenic mice that overexpress cell surface beta1,4-galactosyltransferase Development, January 9, 1996; 122(9): 2859 - 2872. [Abstract] [PDF]

				X Gong, D. Dubois, D. Miller, and B. Shur Activation of a G protein complex by aggregation of beta-1,4-galactosyltransferase on the surface of sperm Science, September 22, 1995; 269(5231): 1718 - 1721. [Abstract] [PDF]