The Lec4A CHO glycosylation mutant arises from miscompartmentalization of a Golgi glycosyltransferase

doi:10.1083/jcb.109.5.2089

This Article

	Full Text (PDF, 943K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Chaney, W.
	Articles by Stanley, P.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Chaney, W.
	Articles by Stanley, P.


Social Bookmarking

	What's this?

ARTICLES

The Lec4A CHO glycosylation mutant arises from miscompartmentalization of a Golgi glycosyltransferase

W Chaney, S Sundaram, N Friedman and P Stanley
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461.

Two CHO glycosylation mutants that were previously shown to lack N- linked carbohydrates with GlcNAc beta 1,6Man alpha 1,6 branches, and to belong to the same genetic complementation group, are shown here to differ in the activity of N-acetylglucosaminyltransferase V (GlcNAc-TV) (UDP-GlcNA: alpha 1,6mannose beta-N-acetylglucosaminyltransferase V). One mutant, Lec4, has no detectable GlcNAc-TV activity whereas the other, now termed Lec4A, has activity equivalent to that of parental CHO in detergent cell extracts. However, Lec4A GlcNAc-TV can be distinguished from CHO GlcNAc-TV on the basis of its increased sensitivity to heat inactivation and its altered subcellular compartmentalization. Sucrose density gradient fractionation shows that the major portion of GlcNAc-TV from Lec4A cells cofractionates with membranes of the ER instead of Golgi membranes where GlcNAc-TV is localized in parental CHO cells. Other experiments show that Lec4A GlcNAc-TV is not concentrated in lysosomes, or in a post-Golgi compartment, or at the cell surface. The altered localization in Lec4A cells is specific for GlcNAc-TV because two other Lec4A Golgi transferases cofractionate at the density of Golgi membranes. The combined data suggest that both lec4 and lec4A mutations affect the structural gene for GlcNAc-TV, causing either the loss of GlcNAc-TV activity (lec4) or its miscompartmentalization (lec4A). The identification of the Lec4A defect indicates that appropriate screening of different glycosylation-defective mutants should enable the isolation of other mammalian cell trafficking mutants.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Cha, S.-K., Hu, M.-C., Kurosu, H., Kuro-o, M., Moe, O., Huang, C.-L. (2009). Regulation of Renal Outer Medullary Potassium Channel and Renal K+ Excretion by Klotho. Mol. Pharmacol. 76: 38-46 [Abstract] [Full Text]
Lau, K. S, Dennis, J. W (2008). N-Glycans in cancer progression. Glycobiology 18: 750-760 [Abstract] [Full Text]
Cha, S.-K., Ortega, B., Kurosu, H., Rosenblatt, K. P., Kuro-o, M., Huang, C.-L. (2008). Removal of sialic acid involving Klotho causes cell-surface retention of TRPV5 channel via binding to galectin-1. Proc. Natl. Acad. Sci. USA 105: 9805-9810 [Abstract] [Full Text]
Patnaik, S. K., Potvin, B., Carlsson, S., Sturm, D., Leffler, H., Stanley, P. (2006). Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells. Glycobiology 16: 305-317 [Abstract] [Full Text]
Patnaik, S. K., Stanley, P. (2005). Mouse Large Can Modify Complex N- and Mucin O-Glycans on {alpha}-Dystroglycan to Induce Laminin Binding. J. Biol. Chem. 280: 20851-20859 [Abstract] [Full Text]
Guo, H.-B., Lee, I., Bryan, B. T., Pierce, M. (2005). Deletion of Mouse Embryo Fibroblast N-Acetylglucosaminyltransferase V Stimulates {alpha}5{beta}1 Integrin Expression Mediated by the Protein Kinase C Signaling Pathway. J. Biol. Chem. 280: 8332-8342 [Abstract] [Full Text]
Ozaslan, D., Wang, S., Ahmed, B. A., Kocabas, A. M., McCastlain, J. C., Bene, A., Kilic, F. (2003). Glycosyl Modification Facilitates Homo- and Hetero-oligomerization of the Serotonin Transporter: A SPECIFIC ROLE FOR SIALIC ACID RESIDUES. J. Biol. Chem. 278: 43991-44000 [Abstract] [Full Text]
Harley, C. A., Dasgupta, A., Wilson, D. W. (2001). Characterization of Herpes Simplex Virus-Containing Organelles by Subcellular Fractionation: Role for Organelle Acidification in Assembly of Infectious Particles. J. Virol. 75: 1236-1251 [Abstract] [Full Text]
Najjar, S. M., Broyart, J.-P., Hampp, L. T., Gray, G. M. (2001). Intestinal aminooligopeptidase in diabetic BioBreed rat: altered posttranslational processing and trafficking. Am. J. Physiol. Gastrointest. Liver Physiol. 280: G104-G112 [Abstract] [Full Text]
Raju, T. S., Stanley, P. (1998). Gain-of-function Chinese Hamster Ovary Mutants LEC18 and LEC14 Each Express a Novel N-Acetylglucosaminyltransferase Activity. J. Biol. Chem. 273: 14090-14098 [Abstract] [Full Text]
Doucey, M.-A., Hess, D., Cacan, R., Hofsteenge, J. (1998). Protein C-Mannosylation Is Enzyme-catalysed and Uses Dolichyl-Phosphate-Mannose as a Precursor. Mol. Biol. Cell 9: 291-300 [Abstract] [Full Text]
Muller, L., Zhu, X., Lindberg, I. (1997). Mechanism of the Facilitation of PC2 Maturation by 7B2: Involvement in ProPC2 Transport and Activation but Not Folding. JCB 139: 625-638 [Abstract] [Full Text]
Whitehouse, C., Burchell, J., Gschmeissner, S., Brockhausen, I., Lloyd, K. O., Taylor-Papadimitriou, J. (1997). A Transfected Sialyltransferase That Is Elevated in Breast Cancer and Localizes to the medial/trans-Golgi Apparatus Inhibits the Development of core-2-based O-Glycans. JCB 137: 1229-1241 [Abstract] [Full Text]
Weinstein, J., Sundaram, S., Wang, X., Delgado, D., Basu, R., Stanley, P. (1996). A Point Mutation Causes Mistargeting of Golgi GlcNAc-TV in the Lec4A Chinese Hamster Ovary Glycosylation Mutant. J. Biol. Chem. 271: 27462-27469 [Abstract] [Full Text]
Lee, J., Sundaram, S., Shaper, N. L., Raju, T. S., Stanley, P. (2001). Chinese Hamster Ovary (CHO) Cells May Express Six beta 4-Galactosyltransferases (beta 4GalTs). CONSEQUENCES OF THE LOSS OF FUNCTIONAL beta 4GalT-1, beta 4GalT-6, OR BOTH IN CHO GLYCOSYLATION MUTANTS. J. Biol. Chem. 276: 13924-13934 [Abstract] [Full Text]