2 Unité de Glycobiologie et Signalisation Cellulaire, INSERM U504, GDR CNRS 2590, Université de Paris Sud XI, 16 Ave Paul Vaillant-Couturier, 94807 Villejuif Cedex France, and 3 CERMAV-CNRS, BP 53, 38041 Grenoble Cedex 9, France
Received on July 9, 2003; accepted on August 14, 2003
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Abstract |
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Key words:
2-fucosyltransferase
/
6-fucosyltransferase
/
conserved peptide motifs
/
phylogeny
/
protein-O-fucosyltransferase
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Introduction |
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The FUT1, FUT2, and Sec1 have a high degree of similarity and constitute a single family of 2-FUTs. The
3- and
4-FUTs are different from the
2-FUTs and are considered as a single family of
3/4-FUTs because they also have a high degree of similarity. Furthermore, in some cases it has been demonstrated that both
1,3 and
1,4 activities can be carried on by the same enzyme (FUT3 or FUT5) (Costache et al., 1997
), and the change of a single amino acid can modify their relative
1,3- or
1,4-activities (Dupuy et al., 1999
, 2002
). The
2- and
6-FUTs share a lower degree of similarity between them, and only the highly sensitive bidimensional hydrophobic cluster analysis (HCA) revealed the presence of three common conserved peptide motifs (I, II, and III), which constitute a signature for these enzymes (Breton et al., 1998
; Chazalet et al., 2001
). This has led us to consider
2- and
6-FUTs as part of the same superfamily originated from a common ancestor by duplication followed by divergent evolution (Oriol et al., 1999
).
When originally described, the first family of POFUT was thought to be independent from all the other known FUTs (Wang et al., 2001), but we show now that there are two families of POFUTs, POFUT1 and POFUT2, which are related to
2- and
6-FUTs.
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Results and discussion |
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Conserved peptide motifs
The HCA of the human POFUT1 and POFUT2 allowed us to identify the same three conserved peptide motifs (I, II, and III) of 2- and
6-FUTs in the POFUT (Figure 2), suggesting that
2-,
6- and protein-O-FUTs belong to the same superfamily.
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At the level of the peptide motifs, the most closely related enzymes are the two 2-FUTs, FUT1 and FUT2, that share 53 out of the 56 conserved positions (95%). The two POFUTs share 28 conserved positions (50%). FUT1 and FUT8 share 25 conserved positions (45%). FUT1 and POFUT1 share 23 conserved positions (41%), and finally FUT1 and POFUT2 are the less similar with only 21 shared conserved positions (38%). FUT8 and POFUT2 share 28 conserved positions (50%) suggesting that FUT8 has more similarity with POFUTs than with
2-FUTs (45%) (Figure 1). This is in good agreement with the shapes of the hydrophobic clusters detected in the human conserved motifs by HCA (Figure 2).
Phylogeny
The phylogenetic analysis was carried on the selected 54 amino acid positions of the block of 56 positions, constituted by the juxtaposition of the 3 conserved peptide motifs shown in Figure 1. The expected four families appear clearly, and the roots of each of these four families have highly significant bootstrap values (8799%) (Figure 3), illustrating that in spite of the presence of numerous conserved positions, there are enough differences among the conserved peptide motifs to clearly cluster the four enzyme families separately.
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Recently two new putative human FUTs with the common conserved peptide motifs of 3-FUTs have been described and called FUT10 (AJ512465) and FUT11 (BC036037) (Roos et al., 2002
). Although they are not yet fully characterized, these last two enzymes have been added to the GDB list of FUTs that now has 11 members, from FUT1 to FUT11. Following the same line of thought, human POFUT1 and POFUT2 should receive the following numbers in this series of human FUTs (FUT12 and FUT13, respectively), because the phylogenetic evidence suggests that they have a common genetic origin, they belong to the same superfamily as the
2- and
6-FUTs, and they use the same GDP-Fuc donor substrate as does every other FUT (Figure 3).
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Materials and methods |
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The conserved peptide motifs I, II, and III of human FUT1, FUT2, FUT8, POFUT1, and POFUT2, were visually identified by HCA (Lemesle-Varloot et al., 1990) (http://smi.snv.jussieu.fr/hca/hca-form.html) (Figure 2). Then the amino acid conserved positions within the peptide motifs of all the enzymes were identified by CLUSTAL W alignment (Thomson et al., 1994
).
We have previously identified 18 putative 2-FUTs in C. elegans (Oriol et al., 1999
); one of them (P91200) has been certified to have
2-FUT activity (Zheng et al., 2002
). We selected only two of the C. elegans
2-FUTs, because all the C. elegans
2-FUTs appear in a single cluster, branching out from the main evolutionary path, before the separation of the vertebrate FUT1 and FUT2 enzymes (Oriol et al., 1999
). In addition, the X. laevis
2-FUT and four species with known FUT1 and FUT2 enzymes (human, murine, porcine, and bovine) were added to complete the
2-FUT family. The
2-FUTs have not been detected among insects (Roos et al., 2002
), urochordates, or birds (this study).
Phylogeny was made with Neighbor Joining (PHYLOWIN package) (Galtier et al., 1996), with G-BLOCKS (Castresana, 2000
), and observed distances. Five hundred sets of data were used for bootstrap calculations (Figure 3).
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Acknowledgements |
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Footnotes |
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Abbreviations |
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References |
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