JBC Transcription and Nuclear Factor Monoclonals

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M102555200 on June 21, 2001

J. Biol. Chem., Vol. 276, Issue 36, 33952-33963, September 7, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/36/33952    most recent
M102555200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van der Wel, H.
Right arrow Articles by West, C. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van der Wel, H.
Right arrow Articles by West, C. M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

A Non-Golgi alpha 1,2-Fucosyltransferase That Modifies Skp1 in the Cytoplasm of Dictyostelium*

Hanke van der WelDagger , Howard R. Morris§, Maria Panico§, Thanai Paxton§, Simon J. North§, Anne Dell§, J. Michael ThomsonDagger , and Christopher M. WestDagger

From the Dagger  Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, Florida 32610-0235 and the § Department of Biochemistry, Imperial College, London SW7 2AY United Kingdom

Skp1 is a subunit of the SCF-E3 ubiquitin ligase that targets cell cycle and other regulatory factors for degradation. In Dictyostelium, Skp1 is modified by a pentasaccharide containing the type 1 blood group H trisaccharide at its core. To address how the third sugar, fucose alpha 1,2-linked to galactose, is attached, a proteomics strategy was applied to determine the primary structure of FT85, previously shown to copurify with the GDP-Fuc:Skp1 alpha 1,2-fucosyltransferase. Tryptic-generated peptides of FT85 were sequenced de novo using Q-TOF tandem mass spectrometry. Degenerate primers were used to amplify FT85 genomic DNA, which was further extended by a novel linker polymerase chain reaction method to yield an intronless open reading frame of 768 amino acids. Disruption of the FT85 gene by homologous recombination resulted in viable cells, which had altered light scattering properties as revealed by flow cytometry. FT85 was necessary and sufficient for Skp1 fucosylation, based on biochemical analysis of FT85 mutant cells and Escherichia coli that express FT85 recombinantly. FT85 lacks sequence motifs that characterize all other known alpha 1,2-fucosyltransferases and lacks the signal-anchor sequence that targets them to the secretory pathway. The C-terminal region of FT85 harbors motifs found in inverting Family 2 glycosyltransferase domains, and its expression in FT85 mutant cells restores fucosyltransferase activity toward a simple disaccharide substrate. Whereas most prokaryote and eukaryote Family 2 glycosyltransferases are membrane-bound and oriented toward the cytoplasm where they glycosylate lipid-linked or polysaccharide precursors prior to membrane translocation, the soluble, eukaryotic Skp1-fucosyltransferase modifies a protein that resides in the cytoplasm and nucleus.

* This work was supported by National Institutes of Health Grant GM-37539 (to C. M. W.) and grants from the Wellcome Trust and BBSRC (to H. R. M. and A. D.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF279134.

To whom correspondence should be addressed: Dept. of Anatomy & Cell Biology, University of Florida College of Medicine, 1600 SW Archer Road, Rm. B1-3, Gainesville, FL 32610-0235. Tel.: 352-392-3329; Fax: 352-392-3305; E-mail: westcm@college.med.ufl.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 GlycobiologyHome page
Q. Zheng, I. Van Die, and R. D Cummings
A novel {alpha}1,2-fucosyltransferase (CE2FT-2) in Caenorhabditis elegans generates H-type 3 glycan structures
Glycobiology, April 1, 2008; 18(4): 290 - 302.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
T. Metcalf, H. van der Wel, R. Escalante, L. Sastre, and C. M. West
Role of SP65 in Assembly of the Dictyostelium discoideum Spore Coat
Eukaryot. Cell, July 1, 2007; 6(7): 1137 - 1149.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
B. Ma, J. L. Simala-Grant, and D. E. Taylor
Fucosylation in prokaryotes and eukaryotes
Glycobiology, December 1, 2006; 16(12): 158R - 184R.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Ercan, M. Panico, M. Sutton-Smith, A. Dell, H. R. Morris, K. L. Matta, D. F. Gay, and C. M. West
Molecular Characterization of a Novel UDP-galactose:Fucoside {alpha}3-Galactosyltransferase That Modifies Skp1 in the Cytoplasm of Dictyostelium
J. Biol. Chem., May 5, 2006; 281(18): 12713 - 12721.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. van der Wel, A. Ercan, and C. M. West
The Skp1 Prolyl Hydroxylase from Dictyostelium Is Related to the Hypoxia-inducible Factor-{alpha} Class of Animal Prolyl 4-Hydroxylases
J. Biol. Chem., April 15, 2005; 280(15): 14645 - 14655.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Ketcham, F. Wang, S. Z. Fisher, A. Ercan, H. van der Wel, R. D. Locke, k. Sirajud-Doulah, K. L. Matta, and C. M. West
Specificity of a Soluble UDP-Galactose:Fucoside {alpha}1,3-Galactosyltransferase That Modifies the Cytoplasmic Glycoprotein Skp1 in Dictyostelium
J. Biol. Chem., July 9, 2004; 279(28): 29050 - 29059.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. M. Moody, S. J. North, B. Reinhold, S. J. Van Dyken, M. E. Rogers, M. Panico, A. Dell, H. R. Morris, J. D. Marth, and E. L. Reinherz
Sialic Acid Capping of CD8beta Core 1-O-Glycans Controls Thymocyte-Major Histocompatibility Complex Class I Interaction
J. Biol. Chem., February 21, 2003; 278(9): 7240 - 7246.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. van der Wel, H. R. Morris, M. Panico, T. Paxton, A. Dell, L. Kaplan, and C. M. West
Molecular Cloning and Expression of a UDP-N-acetylglucosamine (GlcNAc):Hydroxyproline Polypeptide GlcNAc-transferase That Modifies Skp1 in the Cytoplasm of Dictyostelium
J. Biol. Chem., November 22, 2002; 277(48): 46328 - 46337.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. van der Wel, S. Z. Fisher, and C. M. West
A Bifunctional Diglycosyltransferase Forms the Fucalpha 1,2Galbeta 1,3-Disaccharide on Skp1 in the Cytoplasm of Dictyostelium
J. Biol. Chem., November 22, 2002; 277(48): 46527 - 46534.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Q. Zheng, I. Van Die, and R. D. Cummings
Molecular Cloning and Characterization of a Novel alpha 1,2-Fucosyltransferase (CE2FT-1) from Caenorhabditis elegans
J. Biol. Chem., October 11, 2002; 277(42): 39823 - 39832.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
C. M. West, H. van der Wel, and E. A. Gaucher
Complex glycosylation of Skp1 in Dictyostelium: implications for the modification of other eukaryotic cytoplasmic and nuclear proteins
Glycobiology, February 1, 2002; 12(2): 17R - 27R.
[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 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.