Identification of the UDP-glucose-4-epimerase required for galactofuranose biosynthesis and galactose metabolism in A. niger .
Autor: | Park J; Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden University, Sylviusweg 72, Leiden, 2333 BE The Netherlands., Tefsen B; Department of Molecular Cell Biology and Immunology, VU University Medical Center, van den Boechorststraat 7, Amsterdam, 1081 BT The Netherlands.; Present Address: Department of Biological Sciences, Xi'an Jiaotong Liverpool University, 111 Ren Ai Road, Dushu Lake Higher Education Town, Suzhou Industrial Park, Suzhou, 215123 Jiangsu, China., Arentshorst M; Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden University, Sylviusweg 72, Leiden, 2333 BE The Netherlands., Lagendijk E; Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden University, Sylviusweg 72, Leiden, 2333 BE The Netherlands., van den Hondel CA; Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden University, Sylviusweg 72, Leiden, 2333 BE The Netherlands., van Die I; Department of Molecular Cell Biology and Immunology, VU University Medical Center, van den Boechorststraat 7, Amsterdam, 1081 BT The Netherlands., Ram AF; Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden University, Sylviusweg 72, Leiden, 2333 BE The Netherlands. |
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Jazyk: | angličtina |
Zdroj: | Fungal biology and biotechnology [Fungal Biol Biotechnol] 2014 Oct 14; Vol. 1, pp. 6. Date of Electronic Publication: 2014 Oct 14 (Print Publication: 2014). |
DOI: | 10.1186/s40694-014-0006-7 |
Abstrakt: | Background: Galactofuranose (Gal f )-containing glycoconjugates are important to secure the integrity of the cell wall of filamentous fungi. Mutations that prevent the biosynthesis of Gal f -containing molecules compromise cell wall integrity. In response to cell wall weakening, the cell wall integrity (CWI)-pathway is activated to reinforce the strength of the cell wall. Activation of CWI-pathway in Aspergillus niger is characterized by the specific induction of the agsA gene, which encodes a cell wall α-glucan synthase. Results: In this study, we screened a collection of cell wall mutants with an induced expression of agsA for defects in Gal f biosynthesis using a with anti-Gal f antibody (L10). From this collection of mutants, we previously identified mutants in the UDP-galactopyranose mutase encoding gene ( ugmA ). Here, we have identified six additional UDP-galactopyranose mutase ( ugmA ) mutants and one mutant (named mutant #41) in an additional complementation group that displayed strongly reduced Gal f -levels in the cell wall. By using a whole genome sequencing approach, 21 SNPs in coding regions were identified between mutant #41 and its parental strain which changed the amino acid sequence of the encoded proteins. One of these mutations was in gene An14g03820, which codes for a putative UDP-glucose-4-epimerase (UgeA). The A to G mutation in this gene causes an amino acid change of Asn to Asp at position 191 in the UgeA protein. Targeted deletion of ugeA resulted in an even more severe reduction of Gal f in N-linked glucans, indicating that the UgeA protein in mutant #41 is partially active. The ugeA gene is also required for growth on galactose despite the presence of two UgeA homologs in the A. niger genome. Conclusion: By using a classical mutant screen and whole genome sequencing of a new Gal f -deficient mutant, the UDP-glucose-4-epimerase gene ( ugeA ) has been identified. UgeA is required for the biosynthesis of Gal f as well as for galactose metabolism in Aspergillus niger . |
Databáze: | MEDLINE |
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