*Laboratoire de Glycobiologie et Biotechnologie, EA 3176, Institut des Sciences de la Vie et de la Santé, Faculté des Sciences et Techniques, Limoges Cedex, France;
Glycobiologie, INSERM U504, Université Paris XI, Villejuif Cedex, France;
Laboratoire d'Immunogénétique Moléculaire, Hôpital Purpan, Toulouse, France;
Unité de Génétique Moléculaire Animale, UMR-INRA 1061, Institut des Sciences de la Vie et de la Santé, Faculté des Sciences et Techniques, Limoges Cedex, France
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
Considering that the change of type 2 toward type 1 substrate specificity during mammalian evolution might have been of high significance for primate lineage, we analyzed the molecular status of Lewis-like genes belonging to the three main primate groups that have not been studied to date, prosimians (Eulemur fulvus and Varecia variegata), New World monkeys (Saimiri sciureus), and Cercopitecoides (Macaca mulatta). We show that the 1,4 fucose transfer is not restricted to hominoids and that this activity might have appeared early in the primate lineage. Also, characterization of these new genes provide new insights into the chronology of the duplication events at the time of origin of the human Lewis gene cluster.
![]() |
Materials and Methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Southern Blot Analysis
Human and primate genomic DNA were prepared from blood samples (QIAmp Blood Kit, QIAGEN Inc., Hilden, Germany). Ten micrograms was subjected to digestion with EcoRI and BamHI endonucleases, and the fragments were separated in 1% (w/v) agarose gel. DNA was depurinated for 20 min with 0.25 N HCl, denaturated for 30 min with 0.4 N NaOH, and transferred onto a Hybond-N+ membrane (Amersham Pharmacia Biotech, Europe GmbH, Orsay, France). A 402-bp human FUT3 catalytic domain probe was generated by PCR using Ps5 and Pas4 primers (table 1
). Twenty-five nanograms was labeled with [-32P]dCTP by random priming (GIBCO BRL, Pontoise, France). Hybridization was carried out for 14 h at 65°C in a buffer containing 10% (w/v) dextran sulfate, 1% (w/v) SDS, 0.5 M NaCl, and 100 µg of sheared salmon sperm DNA. Blot was washed three times at 42°C for 10 min each with 2 x SSC, 2 x SSC0.1% SDS, and 1 x SSC0.1% SDS and then analyzed by phosphorimaging.
|
Molecular and Phylogenetic Analyses
Nucleotide and amino acid sequences of characterized 3- and
3/4-fucosyltransferases were retrieved from GenBankTM database by BLAST (Altschul et al. 1990
), using the sequences determined in this study. Their accession numbers are Homo sapiens FUT3 (X53578), FUT5 (M81485), and FUT6 (L01698); Pan troglodytes FUT3 (Y14033), FUT5 (Y14034), and FUT6 (Y14035); Bos taurus FUTb (X87810). Alignments were performed with ClustalW version 1.7 (Thompson, Higgins, and Gibson 1994
) and refined with further manual adjustments using the ED program (Philippe 1993
) of the MUST 2000 package (http://sorex.snv.jussieu.fr/must2000.html). Positions that could not be confidently aligned, i.e., the stem indels, and missing data for lemur sequences were removed from the phylogenetic analyses. Phylogenetic trees were constructed with maximum-likelihood (ML), maximum-parsimony (MP), and distance-based (DB) methods with the programs NUCML from MOLPHY version 2.2 (Adachi and Hasegawa 1996
), and DNAPARS and FITCH from PHYLIP version 3.5c (Felsenstein 1989
), respectively. The distances were computed with the two-parameter substitution model of Kimura (1981
). ML trees were obtained by the quick-add operational taxonomic units (OTUs) search with the Hasegawa-Kishino-Yano model of nucleic acid substitution (Hasegawa, Kishino, and Yano 1985
) retaining the 1,000 top ranking trees. A single most parsimonious tree (length = 415) was obtained, and the length (L) of the ML tree was ln L = -2650.94. Bootstrap proportions (Felsenstein 1985
) were calculated by the analysis of 1,000 resampled data sets for MP and DB methods. For ML, bootstrap proportions were computed by using the RELL method (Kishino, Miyata, and Hasegawa 1990
).
Site-directed Mutagenesis
The mammalian expression vector pcDNA1/Amp (InVitrogen) containing the coding sequence of the rhesus macaque FUT5 was directly used for PCR-based mutagenesis. The Arg124Trp mutation was performed with the QuickChange Kit (Stratagene, La Jolla, Calif.). The reaction was carried out with 50 ng of vector, 125 ng of sense primer 5'-GACGCGGTCATCGTGCACCACTGGGATATCATGTAC-3', 125 ng of antisense primer 5'-GTACATGATATCCCAGTGGTGCACGATGACCGCGTC-3', and 2.5 U of Pfu DNA polymerase (Stratagene). After 30 s of denaturation at 95°C, the reaction was cycled for 18 rounds (30 s at 95°C, 1 min at 55°C, and 15 min at 68°C). Parental DNA was removed by DpnI endonuclease treatment. Escherichia coli DH5
strain was directly transformed with the amplified vector. The mutation was verified by sequencing as described previously.
Expression of Mutated Enzymes
Highly pure recombinant plasmids were obtained by anion exchange chromatography (plasmid midi Kit, QIAGEN Inc.). They were used to transiently transfect COS-7 cells using SuperFect Transfection Reagent (QIAGEN Inc.). After 48 h of transfection, proteins were extracted in a lysis buffer (1% [v/v] Triton X100, 10 mM sodium cacodylate (pH 6), 20% [v/v] glycerol, and 1 mM DTT) for 2 h at 4°C. Cellular fragments were eliminated by centrifugation at 4°C (12,000 rpm for 10 min). Protein content was determined with bovine serum albumin as a standard (Bio-Rad, Hercules Calif.) (Bradford 1976
).
Western Blot Analysis
Anti-human FUT3 antibodies were obtained by immunization of rabbits with a truncated form of FUT3 (Pro45-Thr361). This shortened enzyme was produced in E. coli BL21(DE3) cytoplasm associated with a NH2-(His)6-Tag and was purified by affinity chromatography on Ni-NTA-agarose (QIAGEN Inc.). IgG was purified from serum using protein G (HiTrap Protein G, Amersham Pharmacia Biotech. Europe GmbH, Orsay, France). Fifty micrograms of soluble proteins extracted from COS-7 cells was boiled for 3 min after addition of ß-mercaptoethanol (5% v/v) and bromophenol blue (0.02% w/v). Electrophoresis was carried out in Tris/Tricine10.5% SDSpolyacrylamide gel. Separated proteins were then electrotransferred onto a nitrocellulose membrane (Schleicher & Schuell, Dassel, Germany). The immunoblot was processed by chemiluminescence detection (Chemiluminescence Blotting Substrate [POD], Roche Molecular Biochemicals, Mannheim, Germany). The blot was first incubated with rabbit anti-human FUT3 antibodies (4 µg/ml) and then with the secondary antibody, a pig anti-rabbit IgG conjugated to horseradish peroxidase (dilution 1:1,000) (Dako, Denmark).
Fucosyltransferase Assays
Fucosyltransferase assays were performed in a mixture (60 µl) containing 25 mM sodium cacodylate (pH 6.5), 5 mM ATP, 20 mM MnCl2, 10 mM -l-fucose, 3 µM GDP-[14C]-fucose (310 mCi/mmol; Amersham Pharmacia Biotech., U.K.), and 20 µg of crude protein extracts from transfected COS-7 cells. The mixture was incubated for 1 or 3 h at 37°C depending on the activity levels. Acceptor substrate concentrations were 0.1 mM. The reaction was stopped by the addition of 3 ml of cold water. The reaction mixture was then applied to a conditioned Sep-Pack C18 reverse chromatography cartridge (Waters Millipore, Bedford, Mass.) attached to a 10-ml syringe. Unreacted GDP-[14C]-fucose was washed off with 15 ml of water. The radiolabeled reaction product was eluted with two times 5 ml of ethanol, collected directly into scintillation vials and counted with two volumes of Biodegradable Counting Scintillant (Amersham Pharmacia Biotech.) in a liquid scintillation beta counter (liquid scintillation analyzer, Tri-Carb-2100TR, Packard, USA).
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Three fragments were detected in human (fig. 2
) corresponding to the three Lewis genes (Weston et al. 1992b
). A similar pattern was also observed in rhesus macaque, which could indicate that genes orthologous to the three human Lewis ones are present. An additional weaker band of high molecular size was detected and might result from a partial genomic DNA digestion. Interestingly, in the case of ruffed lemur and squirrel monkey, only two fragments were labeled (fig. 2
), irrespective of the endonucleases used (data not shown).
|
Comparison of Lewis-like Peptide Sequences
The new primate peptide sequences were aligned with human, chimpanzee, and bovine Lewis enzymes (fig. 3
). The motifs I and II, which characterize 3/4-fucosyltransferases, were found in all sequences except in FUT sq2. In this sequence the 17-bp deletion beside the His116 codon induces the truncation of the acceptor-binding motif and the loss of motifs I and II. By a virtual deletion of the nucleotide at position 349, a putative FUT sq2 sequence is obtained, which encodes a typical
3/4-fucosyltransferase with only a deletion of six amino acids in the acceptor-binding motif (FUT sq2 frame 2, fig. 3
). Therefore, FUT sq2 gene might be a recently shaped pseudogene. All the other new
3/4-fucosyltransferase sequences possess the highly conserved acceptor-binding motif. The two conserved N-glycosylated sites of human Lewis enzymes (Asn154 and Asn185 in human FUT3, Christensen et al. 2000
) are also found in the new primate enzymes (fig. 3
). Likewise, the four Cys residues (Cys81-Cys338 and Cys91-Cys341) involved in disulfide bonds of human FUT3 (Holmes et al. 2000
) are conserved at the corresponding positions. In the variable domain, upstream of the acceptor-binding motif, a conserved region of nine amino acids named motif III was identified. This motif slightly differs from the region III described by Trottein et al. (2000)
.
|
Both squirrel monkey sequences are distinguished by a common gap of four amino acids from position 45 to 48. In some respects, the FUT sq1 is similar to FUT5 rh. It possesses the conserved "HHRD" acceptor-binding motif and the two characteristic residues of FUT5 enzymes (Pro146 and Thr147). Compared with FUT sq1, FUT sq2 sequence has two additional deletions leading to the absence of three amino acids at the N-terminal end and to a truncated protein (see previously). Thus, its assignment to one orthology class remains tricky. The partial peptide sequences of lemurs show nine additional amino acids (residues 6068) in the stem domain (fig. 3 ). The "HHWD" acceptor-binding motif is present in FUT va1 and FUT eu1 and "HHRD" in FUT va2 and FUT eu2. These enzymes cannot really be assigned to one of the Lewis orthology classes. Nevertheless, they do not possess any conserved residues characteristic of the FUT6 class.
Phylogenetic Analyses of Lewis-like Sequences
Because of the truncated lemur sequences and the suppression of the stem indels for phylogenetic analyses, only 459 nucleic acid positions have been analyzed; 247 positions correspond to variable sites and 149 are informative for parsimony. Considering that Artiodactyls constitute one of the closest sister group of primates, the bovine sequence was chosen as the outgroup for phylogenetic reconstructions. Furthermore, the ancestor of futb sequence is supposed to be orthologous to the ancestor of primate Lewis genes (Oulmouden et al. 1997
). Phylogenetic analyses were carried out using ML, MP, and DB methods. The three analyses showed the same dichotomy with the prosimian sequences always separated from all the other primate sequences (fig. 4
). FUT va1 and FUT eu1 are grouped, with bootstrap values ranging from 59% to 88%. The squirrel monkey fucosyltransferase sequences form a monophyletic group strongly supported by whatever method was used. The three classes of orthology, FUT3, FUT5, and FUT6, are always recovered with bootstrap values ranging from 98% to 100%. Within the three sequences identified in the rhesus macaque, FUT3 rh and FUT5 rh consistently cluster with their hominoid counterparts, always emerging as their sister group. The third rhesus sequence, FUT3' rh, is related to the FUT3 orthology class, and their clustering is supported by high bootstrap values, 88%97%. The relative branching order of the three orthology classes and the New World monkey group is not resolved (bootstrap values between 15% and 48%). However, analyses performed without lemur sequences support the fact that the Saimiri sequences are the sister group of FUT5 (data not shown).
|
|
|
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Although the Trp residue of acceptor-binding motif is not sufficient by itself to determine 1,4 fucose transfer (Dupuy et al. 1999
), the presence, in the two lemur species, of a Lewis-like sequence with the "HHWD" acceptor-binding motif typical of
3/4-fucosyltransferases suggests the possible existence of
1,4 fucosylation in pro-simians. Conversely, in New World monkeys the
3/4-fucosyltransferase gene could have evolved into a pseudogene, as illustrated by the FUT sq2, and this may justify the absence of Lea antigens (Moor-Jankowski and Wiener 1968
). Thus, the ability to transfer fucose in
1,4 linkage is not restricted to hominoids and may be generalized at least to catarrhines and possibly to all primates.
The 1,4 fucosylated glycoconjugates are also present in plants. Recently, two
1,4-fucosyltransferase activities were characterized from Vaccinium myrtillus (Palma et al. 2001
) and Arabidopsis thaliana (Léonard, personal communication). Interestingly, plant
4-fucosyltransferase genes (Bakker et al. 2001
) encode proteins with the characteristic motif II and a putative acceptor-binding motif similar to the primate one. These shared motifs suggest a common origin of the genes responsible for
1,4 fucosylation in primates and plants. However, because such activity is not found in other vertebrates, i.e., other mammals belonging to Carnivores, Artiodactyls, Lagomorphs, and Rodents, which are Lea negative (Oriol et al. 1999
), we have to admit that the
1,4 fucosylation has independently appeared twice, in plants and in primates.
In animals the simplest model of Lewis fucosyltransferase gene evolution (fig. 6A
), which explains the emergence of 1,4 fucosylation, would involve two successive duplications of a Lewis ancestral gene, after the great mammalian radiation 80 MYA (Oulmouden et al. 1997
; Wierinckx et al. 1999
). In primates after the first duplication of a gene orthologous to the bovine futb ancestral gene, a second duplication would be at the origin of FUT3 and FUT5, the corresponding enzymes carrying the
1,4 activity (Dupuy et al. 1999
). The presence of two Lewis-like sequences in pro-simians (brown and ruffed lemurs) and in New World monkeys (squirrel monkey) suggests that the first duplication event occurred before the emergence of pro-simians, 70 MYA (fig. 6B
). The three genes identified in rhesus macaque could set the second duplication event before the separation of Cercopitecoides from hominoids 35 MYA and after the emergence of Platyrrhines 47 MYA. However, species-specific independent gene duplications could not be formally ruled out.
|
Interestingly, the 1,4 fucosylation is supported by one gene in Cercopitecoides (FUT3 rh), and potentially in pro-simians, whereas in Hominoids two genes (FUT3 and FUT5) are involved. In our hypothesis, the ancestor of FUT3 enzyme might already have an
1,4 activity, whereas FUT5 would have gained this ability after the emergence of Cercopitecoides. Indeed, the rhesus macaque FUT5 enzyme, unlike chimpanzee and human FUT5, possesses the HHRD acceptor-binding motif and uses exclusively type 2 acceptor substrates. The Arg124
Trp substitution in FUT5 rh, which generates the HHWD acceptor-binding motif characteristic of
3/4-fucosyltransferases, produces an enzyme without type 1 activity and which has lost its type 2 substrate specificity. This result shows that one mutation from Arg to Trp is sufficient to abolish the
1,3 activity, but it is not enough to acquire the
1,4 fucosylation, which probably needs the mutation of other amino acids. The substitution Arg
Trp in the acceptor-binding motif of FUT5 might have occurred only recently in hominoids. Therefore, the
1,4 fucosylation might have appeared twice independently in the primate lineage. Nevertheless, in regard to the rhesus macaque sequences, evolution of Lewis genes seems more complicated than was suggested before and requires new genetic information from other primates.
![]() |
Acknowledgements |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Footnotes |
---|
Abbreviations: Fuc, fucose; FUT, fucosyltransferase; FUT3, FUT3-encoded Lewis 3/4-fucosyltransferase; FUT5, FUT5-encoded
3/4-fucosyltransferase; FUT6, FUT6-encoded plasma
3-fucosyltransferase; FUTb, bovine futb-encoded
3-fucosyltransferase; Gal, galactose; GlcNAc, N-acetylglucosamine; Tricine, N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl] glycine; The Lewis blood group antigens are Lea, Galß1,3(Fuc
1,4)GlcNAc; Leb, Fuc
1,2Galß1,3(Fuc
1,4) GlcNAc; Lex, Galß1,4(Fuc
1,3)GlcNAc; Ley, Fuc
1,2Galß1,4(Fuc
1,3)GlcNAc.
Keywords: acceptor substrate specificity
3/4-fucosyltransferase
evolution
Lewis
phylogeny
primate
Address for correspondence and reprints: Abderrahman Maftah, Laboratoire de Glycobiologie et Biotechnologie, EA 3176, Faculté des Sciences et Techniques, Université de Limoges, 123 Avenue Albert Thomas, 87060 Limoges Cedex, France. maftah{at}unilim.fr
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Adachi J., M. Hasegawa, 1996 MOLPHY version 2.3: programs for molecular phylogenetics based on maximum likelihood Comput. Sci. Monogr 28:1-150
Altschul S. F., W. Gish, W. Miller, E. W. Myers, D. J. Lipman, 1990 Basic local alignment search tool J. Mol. Biol 215:403-410[ISI][Medline]
Bakker H., E. Schijlen, T. de Vries, W. E. C. M. Schiphorst, W. Jordi, A. Lommen, D. Bosch, I. van Die, 2001 Plant members of 1
3/4-fucosyltransferase gene family encode an
1
4-fucosyltransferase, potentially involved in Lewisa biosynthesis, and two core
1
3-fucosyltransferases FEBS Lett 507:307-312[ISI][Medline]
Bradford M. M., 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal. Biochem 72:248-254[ISI][Medline]
Candelier J. J., R. Mollicone, B. Mennesson, P. Couillin, R. Oriol, 2000 Expression of fucosyltransferases in skin, conjunctiva, and cornea during human development Histochem. Cell Biol 114:113-124[ISI][Medline]
Christensen L. L., U. B. Jensen, P. Bross, T. F. Orntoft, 2000 The C-terminal N-glycosylation sites of the human 1,3/4-fucosyltransferase III, -V, and -VI (hFucTIII, -V, and -VI) are necessary for the expression of full enzyme activity Glycobiology 10:931-939
Costache M., P. Apoil, A. Cailleau, A. Elmgren, G. Larson, S. Henry, A. Blancher, D. Iordachescu, R. Oriol, R. Mollicone, 1997a. Evolution of fucosyltransferase genes in vertebrates J. Biol. Chem 272:29721-29728
Costache M., A. Cailleau, P. Fernandez-Mateos, R. Oriol, R. Mollicone, 1997b. Advances in molecular genetics of -2- and
-3/4-fucosyltransferases Transfus. Clin. Biol 4:367-382[ISI][Medline]
Dupuy F., J. M. Petit, R. Mollicone, R. Oriol, R. Julien, A. Maftah, 1999 A single amino acid in the hypervariable stem domain of vertebrate 1,3/1,4-fucosyltransferases determines the type1/type2 transfer: characterization of acceptor substrate specificity of the Lewis enzyme by site-directed mutagenesis J. Biol. Chem 274:12257-12262
Elmgren A., C. Borjeson, R. Mollicone, R. Oriol, A. Fletcher, G. Larson, 2000 Identification of two functionally deficient plasma alpha 3-fucosyltransferase (FUT6) alleles Hum. Mutat 16:473-481[ISI][Medline]
Felsenstein J., 1985 Confidence limits on phylogenies: an approach using the bootstrap Evolution 40:783-791
. 1989 PHYLIP (phylogeny inference package). Version 3.2 Cladistics 5:164-166
Gersten K. M., S. Natsuka, M. Trinchera, B. Petryniak, R. J. Kelly, N. Hiraiwa, N. A. Jenkins, D. J. Gilbert, N. G. Copeland, J. B. Lowe, 1995 Molecular cloning, expression, chromosomal assignment, and tissue-specific expression of a murine -(1,3)-fucosyltransferase locus corresponding to the human ELAM-1 ligand fucosyl transferase J. Biol. Chem 270:25047-25056
Goelz S. E., C. Hession, D. Goff, B. Griffiths, R. Tizard, B. Newman, G. Chi-Rosso, R. Lobb, 1990 ELFT: a gene that directs the expression of an ELAM-1 ligand Cell 63:1349-1356[ISI][Medline]
Hasegawa M., H. Kishino, T. Yano, 1985 Dating of the human-ape splitting by a molecular clock of mitochondrial DNA J. Mol. Evol 22:160-174[ISI][Medline]
Holmes E. H., T. Y. Yen, S. Thomas, R. Joshi, A. Nguyen, T. Long, F. Gallet, A. Maftah, R. Julien, B. A. Macher, 2000 Human 1,3/4 fucosyltransferases: characterization of highly conserved cysteine residues and N-linked glycosylation sites J. Biol. Chem 275:24237-24245
Kaneko M., T. Kudo, H. Iwasaki, et al. (11 co-authors) 1999 1,3-Fucosyltransferase IX (Fuc-TIX) is very highly conserved between human and mouse; molecular cloning, characterization and tissue distribution of human Fuc-TIX FEBS Lett 452:237-242[ISI][Medline]
Kimura M., 1981 A simple method for estimating evolutionary rates of base substitution through comparative studies of nucleotide sequences J. Mol. Evol 16:111-120[ISI]
Kishino H., T. Miyata, M. Hasegawa, 1990 Maximum likelihood inference of protein phylogeny, and the origin of chloroplasts J. Mol. Evol 31:151-160[ISI]
Kukowska-Latallo J. F., R. D. Larsen, R. P. Nair, J. B. Lowe, 1990 A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group alpha(1,3/1,4)fucosyltransferase Genes Dev 4:1288-1303[Abstract]
Legault D. J., R. J. Kelly, Y. Natsuka, J. B. Lowe, 1995 Human (1,3/1,4)-fucosyltransferases discriminate between different oligosaccharide acceptor substrates through a discrete peptide fragment J. Biol. Chem 270:20987-20996
Mollicone R., A. Gibaud, A. Francois, M. Ratcliffe, R. Oriol, 1990 Acceptor specificity and tissue distribution of three human alpha-3-fucosyltransferases Eur. J. Biochem 191:169-176[Abstract]
Mollicone R., I. Reguigne, A. Fletcher, A. Aziz, M. Rustam, B. W. Weston, R. J. Kelly, J. B. Lowe, R. Oriol, 1994a. Molecular basis for plasma (1,3)-fucosyltransferase gene deficiency (FUT6) J. Biol. Chem 269:12662-12671
Mollicone R., I. Reguigne, R. J. Kelly, A. Fletcher, J. Watt, S. Chatfield, A. Aziz, H. S. Cameron, B. W. Weston, J. B. Lowe, 1994b. Molecular basis for Lewis (1,3/1,4)-fucosyltransferase gene deficiency (FUT3) found in Lewis-negative Indonesian pedigrees J. Biol. Chem 269:20987-20994
Moor-Jankowski J., A. S. Wiener, 1968 Blood groups of non-human primates. Summary of the currently available information Primates Med 1:49-67[Medline]
Nguyen A. T., E. H. Holmes, J. M. Whitaker, S. Ho, S. Shetterly, B. A. Macher, 1998 Human 1,3/4-fucosyltransferasesI. Identification of amino acids involved in acceptor substrate binding by site-directed mutagenesis J. Biol. Chem 273:25244-25249
Oriol R., 1995 ABO, Hh, Lewis and secretion. Serology, genetics and tissue distribution Pp. 3773 in J. P. Cartron and P. Rouger, eds. Molecular basis of major blood group antigens. Plenum Press, London
Oriol R., J. J. Candelier, S. Taniguchi, L. Balanzino, L. Peters, M. Niekrasz, C. Hammer, D. K. Cooper, 1999 Major carbohydrate epitopes in tissues of domestic and African wild animals of potential interest for xenotransplantation research Xenotransplantation 6:79-89[ISI][Medline]
Orntoft T. F., E. M. Vestergaard, E. Holmes, et al. (13 co-authors) 1996 Influence of Lewis (1,3/4)-l-fucosyltransferase (FUT3) gene mutations on enzyme activity, erythrocyte phenotyping, and circulating tumor marker sialyl-Lewis a levels J. Biol. Chem 274:32260-32268
Oulmouden A., A. Wierinckx, J. Petit, M. Costache, M. M. Palcic, R. Mollicone, R. Oriol, R. Julien, 1997 Molecular cloning and expression of a bovine (1,3)fucosyltransferase gene homologous to a putative ancestor gene of the human FUT3-FUT5-FUT6 cluster J. Biol. Chem 272:8764-8773
Palma A. S., C. Vila-Verde, A. S. Pires, P. S. Fevereiro, J. Costa, 2001 A novel plant 4-fucosyltransferase (Vaccinium myrtillus L.) synthesises the Lewis(a) adhesion determinant FEBS Lett 499:235-238[ISI][Medline]
Pang H., Y. Koda, M. Soejima, H. Kimura, 1998a. Significance of each of three missense mutations, G484A, G667A, and G808A, present in an inactive allele of the human Lewis gene (FUT3) for (1,3/1,4)fucosyltransferase inactivation Glycoconj. J 15:961-967[ISI][Medline]
Pang H., Y. Koda, M. Soejima, T. Schlaphoff, E. D. du Toit, H. Kimura, 1999 Allelic diversity of the human plasma (1,3)fucosyltransferase gene (FUT6) Ann. Hum. Genet 63:277-284[ISI][Medline]
Pang H., Y. Liu, Y. Koda, M. Soejima, J. Jia, T. Schlaphoff, E. D. Du Toit, H. Kimura, 1998b. Five novel missense mutations of the Lewis gene (FUT3) in African (Xhosa) and Caucasian populations in South Africa Hum. Genet 102:675-680[ISI][Medline]
Philippe H., 1993 MUST, a computer package of management utilities for sequences and trees Nucleic Acids Res 21:5264-5272[Abstract]
Reguigne-Arnould I., P. Couillin, R. Mollicone, S. Faure, A. Fletcher, R. J. Kelly, J. B. Lowe, R. Oriol, 1995 Relative positions of two clusters of human -l-fucosyltransferases in 19q (FUT1-FUT2) and 19p (FUT6-FUT3-FUT5) within the microsatellite genetic map of chromosome 19 Cytogenet. Cell Genet 71:158-162[ISI][Medline]
Sanger F., S. Nicklen, A. R. Coulson, 1977 DNA sequencing with chain-terminating inhibitors Proc. Natl. Acad. Sci. USA 74:5463-5467[Abstract]
Sasaki K., K. Kurata, K. Funayama, M. Nagata, E. Watanabe, S. Ohta, N. Hanai, T. Nishi, 1994 Expression cloning of a novel 1,3-fucosyltransferase that is involved in biosynthesis of the sialyl Lewis x carbohydrate determinants in leukocytes J. Biol. Chem 269:14730-14737
Thompson J. D., D. G. Higgins, T. J. Gibson, 1994 ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice Nucleic Acids Res 22:4673-4680[Abstract]
Trottein F., R. Mollicone, J. Fontaine, R. de Mendonca, F. Piller, R. Pierce, R. Oriol, M. Capron, 2000 Molecular cloning of a putative 3-fucosyltransferase from Schistosoma mansoni Mol. Biochem. Parasitol 107:279-287[ISI][Medline]
Weston B. W., R. P. Nair, R. D. Larsen, J. B. Lowe, 1992a. Isolation of a novel human (1,3)-fucosyltransferase gene and molecular comparison to the human Lewis blood group
(1,3/1,4)-fucosyltransferase gene. Syntenic, homologous, nonallelic genes encoding enzymes with distinct acceptor substrate specificities J. Biol. Chem 267:4152-4160
Weston B. W., P. L. Smith, R. J. Kelly, J. B. Lowe, 1992b. Molecular cloning of a fourth member of a human (1,3)-fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis x, sialyl Lewis x, and difucosyl sialyl Lewis x epitopes J. Biol. Chem 267:24575-24584 [published erratum appears in J. Biol. Chem 1993 268:18398 ]
Wierinckx A., D. Mercier, A. Oulmouden, J. M. Petit, R. Julien, 1999 Complete genomic organization of futb encoding a bovine 3-fucosyltransferase: exons in human orthologous genes emerged from ancestral intronic sequences Mol. Biol. Evol 16:1535-1547[Abstract]
Wietrzyk J., A. Opolski, J. Madej, A. Laskowska, C. Radzikowski, M. Ugorski, 2000 Metastatic potential of human uroepithelial cancer cells is not dependent on their adhesion to E-selectin Anticancer Res 20:913-916[ISI][Medline]
Zhang A., B. Potvin, A. Zaiman, W. Chen, R. Kumar, L. Phillips, P. Stanley, 1999 The gain-of-function Chinese hamster ovary mutant LEC11B expresses one of two Chinese hamster FUT6 genes due to the loss of a negative regulatory factor J. Biol. Chem 274:10439-10450