UDP-N-acetylglucosamine:: α-3-D-mannoside β1,2-N-acetylglucosaminyltransferase I and UDP-N-acetylglucosamine:: α-6-D-mannoside β-1,2-N-acetylglucosaminyltransferase II in Caenorhabditis elegans

被引:29
作者
Chen, SH
Tan, J
Reinhold, VN
Spence, AM
Schachter, H
机构
[1] Hosp Sick Children, Dept Struct Biol & Biochem, Toronto, ON M5G 1X8, Canada
[2] Univ Toronto, Dept Biochem, Toronto, ON M5S 1A8, Canada
[3] Univ New Hampshire, Dept Chem, Durham, NH 03824 USA
[4] Univ Toronto, Dept Mol & Med Genet, Toronto, ON M5S 1A8, Canada
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2002年 / 1573卷 / 03期
基金
加拿大健康研究院;
关键词
complex N-glycans; null mutation; UV-TMP mutagenesis; worm development;
D O I
10.1016/S0304-4165(02)00393-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GnT I) and UDP-N-acetylglucosamine:alpha-6-D-mannoside beta-1,2-N-acetylglucosaminyltransferase II (GnT 11) are key enzymes in the synthesis of Asn-linked hybrid and complex glycans. We have cloned cDNAs from Caenorhabditis elegans for three genes homologous to mammalian GnT I (designated gly-12, gly-13 and gly-14) and one gene homologous to mammalian GnT II. All four cDNAs encode proteins which have the domain structure typical of previously cloned Golgi-type glycosyltransferases and show enzymatic activity (GnT I and GnT 11, respectively) on expression in transgenic worms. We have isolated worm mutants lacking the three GnT I genes by the method of ultraviolet irradiation in the presence of trimethylpsoralen (TMP); null mutants for GnT 11 have not yet been obtained. The gly-12 and gly-14 mutants as well as the gly-14;gly-12 double mutant displayed wild-type phenotypes indicating that neither gly-12 nor gly-14 is necessary for worm development under standard laboratory conditions. This finding and other data indicate that the GLY-13 protein is the major functional GnT I in C elegans. The mutation lacking the gly-13 gene is partially lethal and the few survivors display severe morphological and behavioral defects. We have shown that the observed phenotype co-segregates with the gly-13 deletion in genetic mapping experiments although a second mutation near the gly-13 gene cannot as yet be ruled out. Our data indicate that complex and hybrid N-glycans may play critical roles in the morphogenesis of C elegans, as they have been shown to do in mice and men. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:271 / 279
页数:9
相关论文
共 81 条
[1]   INSECT CELLS CONTAIN AN UNUSUAL, MEMBRANE-BOUND BETA-N-ACETYLGLUCOSAMINIDASE PROBABLY INVOLVED IN THE PROCESSING OF PROTEIN N-GLYCANS [J].
ALTMANN, F ;
SCHWIHLA, H ;
STAUDACHER, E ;
GLOSSL, J ;
MARZ, L .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (29) :17344-17349
[2]   Genetic model organisms in the study of N-glycans [J].
Altmann, F ;
Fabini, G ;
Ahorn, H ;
Wilson, IBH .
BIOCHIMIE, 2001, 83 (08) :703-712
[3]   Insect cells as hosts for the expression of recombinant glycoproteins [J].
Altmann, F ;
Staudacher, E ;
Wilson, IBH ;
März, L .
GLYCOCONJUGATE JOURNAL, 1999, 16 (02) :109-123
[4]   PROCESSING OF ASPARAGINE-LINKED OLIGOSACCHARIDES IN INSECT CELLS - N-ACETYLGLUCOSAMINYLTRANSFERASE I AND II ACTIVITIES IN CULTURED LEPIDOPTERAN CELLS [J].
ALTMANN, F ;
KORNFELD, G ;
DALIK, T ;
STAUDACHER, E ;
GLOSSL, J .
GLYCOBIOLOGY, 1993, 3 (06) :619-625
[5]   On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database [J].
Apweiler, R ;
Hermjakob, H ;
Sharon, N .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1999, 1473 (01) :4-8
[6]   An Arabidopsis thaliana cDNA complements the N-acetylglucosaminyltransferase I deficiency of CHO Lec1 cells [J].
Bakker, H ;
Lommen, A ;
Jordi, W ;
Stiekema, W ;
Bosch, D .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 261 (03) :829-832
[7]   SQV-7, a protein involved in Caenorhabditis elegans epithelial invagination and early embryogenesis, transports UDP-glucuronic acid, UDP-N-acetylgalactosamine, and UDP-galactose [J].
Berninsone, P ;
Hwang, HY ;
Zemtseva, I ;
Horvitz, HR ;
Hirschberg, CB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (07) :3738-3743
[8]   sqv-3 -7, and -8, a set of genes affecting morphogenesis in Caenorhabditis elegans, encode enzymes required for glycosaminoglycan biosynthesis [J].
Bulik, DA ;
Wei, G ;
Toyoda, H ;
Kinoshita-Toyoda, A ;
Waldrip, WR ;
Esko, JD ;
Robbins, PW ;
Selleck, SB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (20) :10838-10843
[9]  
Campbell R, 1997, GLYCOBIOLOGY, V7, P143
[10]  
CHARUK JHM, 1995, EUR J BIOCHEM, V230, P797