Detection and structural mapping of potential fucosyltransferases motif sequences in some insect species

Abstract

Amaç: Fukoziltransferazlar (FucTs), GDP-Fukoz (GDP-Fuc)’tan α-L-fukozun N- ve O-bağlı glikanlara, serbest oligosakkaritlere veya lipitlere ya da doğrudan proteinlere aktarılmasını (fukozilasyon) katalizler. Memelilerde, FUT1–11, POFUT1 (Protein O-fukoziltransferaz 1) ve POFUT2 (Protein O-fukoziltransferaz 2) dahil olmak üzere 13 fukoziltransferaz gen ailesi (FUT) tanımlanmıştır. Literatürde mevcut çalışmalar bulunmasına rağmen, birçok organizmada fukoziltransferazların çeşitliliği hâlâ tam olarak bilinmemektedir. Böceklerde ise fukoziltransferazlar üzerine yapılan çalışmalar oldukça sınırlıdır ve FUT tipleri tür bazında henüz karakterize edilmemiştir. Metot: Genomları bilinen 5 böceğin (Bombyx mori, Callosobruchus maculatus, Galleria mellonella, Manduca sexta, Myzus persicae) veri tabanlarından elde edilen transkriptomlarından (tüm mRNA dizileri) fukoziltransferaz motif dizileri veri tabanları (Clustal Omega, Prosite ve MEME-Suite/FIMO) aracılığıyla tespit edilerek böceklerdeki olası fukoziltransferazlar belirlenmiştir. Daha sonra memelilerdeki fukoziltransferaz enzimlerinin üç boyutlu protein yapıları arasındaki benzerlikler Swiss-Model, AlphaFold ve pyMOL veri tabanları kullanılarak ortaya konulmuştur. Bulgular: Motif ve üç boyutlu yapı karşılaştırma bölgelerine göre, Callosobruchus maculatus türünde 13 fukoziltransferazın tümü bulunurken, Galleria mellonella türünde FucT1 ve FucT9, Manduca sexta türünde FucT1, FucT4, FucT5, FucT8 ve FucT11, Myzus persicae türünde FucT2, FucT3, FucT5, FucT6, FucT7, FucT9, FucT10 ve FucT11, Bombyx mori türünde FucT1, FucT3, FucT5, FucT6 ve FucT9 motifleri karakterize edilmiştir. Tartışma: Bu çalışma, omurgasızlarda fukoziltransferazlar üzerine yapılacak ilerleyen çalışmalar için yol gösterici özellik taşımaktadır. Genomu belirlenen böceklerle çalışıldıkça fukoziltransferaz enzimi için yeni diziler ortaya çıkarılacaktır. Böcekler ve memelilerdeki fukoziltransferaz motiflerinin benzerlikleri, bu genlerin evrimsel açıdan ortolog genler olabileceğine işaret ederken, ayrıca bu konuda gelişimsel ve evrimsel yeni çalışmaların yapılacağının göstergesidir.

Keywords

• Motif
• 3B protein modeli
• Fukoziltransferazlar
• Böcek

Supporting Institution

Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

FYL-2023-27378

Ethical Statement

Bu çalışma için etik onay gerekmemektedir.

Detection and structural mapping of potential fucosyltransferases motif sequences in some insect species

Abstract

Purpose: Fucosyltransferases (FucTs) catalyze the transfer (fucosylation) of α-L-fucose from GDP-Fuc onto N- and O-linked glycans, free oligosaccharides, or lipids or directly onto proteins. In mammals, 13 fucosyltransferase gene families (FUT) have been identified, including FUT 1-11, POFUT1 (Protein O-fucosyltransferase 1) and POFUT2 (Protein O-fucosyltransferase 2) Despite the existing studies in the literature, the diversity of many fucosyltransferases in organisms remains unknown. In insects, there are few studies on fucosyltransferase and FUT types are not characterized by species. Method: Fucosyltransferase motif sequences were determined from the transcriptomes (all mRNA sequences) obtained from the databases of 5 insects (Bombyx mori, Callosobruchus maculatus, Galleria mellonella, Manduca sexta, Myzus persicae) with known genomes through databases, and putative fucosyltransferase sequences in insects were determined with computational analysis (Clustal Omega, Prosite ve MEME-Suite/FIMO). Then, the similarities between the three-dimensional protein structures of the mammalian fucosyltransferase enzymes were revealed using Swiss-Model, AlphaFold and pyMOL databases. Findings: Based on motif and three-dimensional structure comparison regions, all 13 fucosyltransferases were identified in Callosobruchus maculatus, FucT1 and FucT9 in Galleria mellonella, FucT1, FucT4, FucT5, FucT8, and FucT11 in Manduca sexta, FucT2, FucT3, FucT5, FucT6, FucT7, FucT9, FucT10, and FucT11 in Myzus persicae, and FucT1, FucT3, FucT5, FucT6, and FucT9 motifs were characterized in Bombyx mori. Conclusion: This information provides guidance for continuing studies on fucosyltransferases in invertebrates. New sequences for the fucosyltransferase enzyme will be revealed as the genome of determined insects is studied. While the similarities of fucosyltransferase motifs in insects and mammals indicate that they may be orthologous genes in terms of evolution, it is also an indication that new developmental and evolutionary studies will be conducted on this subject.

Keywords

• Motif scanning
• 3D protein model
• fucosyltransferases
• insect

Supporting Institution

Ege University Scientific Research Fund

Project Number

FYL-2023-27378

Ethical Statement

This study does not require ethical approval.

References

1. [1] Oriol, R., Mollicone, R., Cailleau, A., Balanzino, L. and Breton, C. (1999). Divergent evolution of fucosyltransferase genes from vertebrates, invertebrates, and bacteria, Glycobiology 9: 323–334. https://doi.org/10.1093/glycob/9.4.323
2. [2] Wang, Y., Shao, L., Shi, S., Harris, R. J., Spellman, M. W., Stanley, P., & Haltiwanger, S. (2001). Modification of Epidermal Growth Factor-like Repeats with O-Fucose. Journal of Biological Chemistry, 276, 40338–40345. https://doi.org/10.1074/jbc.M107849200
3. [3] Mollicone, R., Moore, S., Bovin, N., Rosasco, M., Candelier, J., Duncker, I., & Oriol, R. (2009). Activity, splice variants, conserved peptide motifs, and phylogeny of two new α1,3-fucosyltransferase families (FUT10 and FUT11). Journal of Biological Chemistry, 284, 4723–4738. https://doi.org/10.1074/jbc.M809312200
4. [4] Shan, M., Yang, D., Dou, H., & Zhang, L. (2019). Fucosylation in cancer biology and its clinical applications. Progress in Molecular Biology and Translational Science, 162, 1877-1173. https://doi.org/10.1016/bs.pmbts.2019.01.002
5. [5] Vries, T., Knegtel, R., Holmes, E., & Macher, B. (2001). Fucosyltransferases: Structure/function studies. Glycobiology, 11, 119-128. https://doi.org/10.1093/glycob/11.10.119R
6. [6] Xu, Z., Vo, L., & Macher, B. A. (1996). Structure-Function Analysis of Human α1,3-Fucosyltransferase. Journal of Biological Chemistry, 271, 8818–8823. https://doi.org/10.1074/jbc.271.15.8818
7. [7] Dupuy, F., Germot, A. M., Oriol, R., Blancher, A., Julien, R., & Maftah, A. (2002). α1,4-Fucosyltransferase activity: A significant function in the primate lineage has appeared twice independently. Molecular Biology Evolution, 19, 815–824. https://doi.org/10.1093/oxfordjournals.molbev.a004138
8. [8] Dupuy, F., Germot, A., Julien, R., & Maftah, A. (2004). Structure/function study of Lewis α3- and a3/4-fucosyltransferases: The a1,4 fucosylation requires an aromatic residue in the acceptor-binding domain. Glycobiology, 14, 347–356. https://doi.org/10.1093/glycob/cwh053

9. [9] Paschinger, K., Staudacher, E., Stemmer, U., Fabini, G., & Wilson, I. B. H. (2005). Fucosyltransferase substrate specificity and the order of fucosylation in invertebrates. Glycobiology, 15, 463–474. https://doi.org/10.1093/glycob/cwi028
10. [10] Ihara, H., Okada, T., & Ikeda, Y. (2014). Cloning, expression and characterization of Bombyx mori α1,6-fucosyltransferase. Biochemical and Biophysical Research Communications, 450, 953–960. https://doi.org/10.1016/j.bbrc.2014.06.087
11. [11] Yan, S., Serna, S., Reichardt, N.C., Paschinger, K., Wilson, I. B. H. (2013). Array-assisted characterization of a fucosyltransferase required for the biosynthesis of complex core modifications of nematode N-Glycans. Journal of Biological Chemistry, 288, 21015–21028. https://doi.org/10.1074/jbc.M113.479147
12. [12] Kurz, S., King, J.G., Dinglasan, R. R., Paschinger, K., Wilson, I. B. H. (2016). The fucomic potential of mosquitoes: Fucosylated N-glycan epitopes and their cognate fucosyltransferases. Insect Biochemistry and Molecular Biology, 68, 52-63. https://doi.org/10.1016/j.ibmb.2015.11.001
13. [13] Roos, C., Kolmer, M., Mattila, P., & Renkonen, R. (2002). Composition of Drosophila melanogaster proteome involved in fucosylated glycan metabolism. The Journal of Biological Chemistry, 277, 3168–3175. https://doi.org/10.1074/jbc.M107927200
14. [14] Breton, C., Oriol, R. & Imberty, A. (1998). Conserved structural features in eukaryotic and prokaryotic fucosyltransferases, Glycobiology, 8, 87–94. https://doi.org/10.1093/glycob/8.1.87
15. [15] Baboval, T., & Smith, F. I. (2002). Comparison of human and mouse Fuc-TX and Fuc-TXI genes, and expression studies in the mouse. Mammalian Genome, 13, 538–541. https://doi.org/10.1007/s00335-001-2152-5.
16. [16] Fabini, G., Freilinger, A., Altmann, F., & Wilson, I. B. H. (2001). Identification of core α1,3-fucosylated glycans and cloning of the requisite fucosyltransferase cDNA from Drosophila melanogaster. Journal of Biological Chemistry, 276, 28058–28067. https://doi.org/10.1074/jbc.M100573200
17. [17] Rendic, D., Klaudiny, J., Stemmer, U., Schmidt, J., Paschinger, K., & Wilson, I. B. H. (2007). Towards abolition of immunogenic structures in insect cells: Characterization of a honey-bee (Apis mellifera) multi-gene family reveals both an allergy-related core α1,3-fucosyltransferase and the first insect Lewis-histo-blood-group-related antigen-synthesizing enzyme. Biochemistry Journal, 402, 105–115. https://doi.org/10.1042/BJ20060964
18. [18] Costache, M., Cailleau, A., Fernandez-Mateos, P., Oriol, R., & Mollicone, R. (1997). Advances in molecular genetics of and (x-3/4-fucosyltransferases). Transfusion Clinique et Biologique, 4, 367–382. https://doi.org/10.1016/S1246-7820(97)80042-0
19. [19] Petit, D., Maftah, A., Julien, R., & Petit, J. M. (2006). En bloc duplications, mutation rates, and densities of amino acid changes clarify the evolution of vertebrate α-1,3/4-fucosyltransferases. Journal of Molecular Evolution, 63, 353–364. https://doi.org/10.1007/s00239-005-0189-x