Functional Analysis of Three Putative Galactofuranosyltransferases with Redundant Functions in Galactofuranosylation in Aspergillus Niger

Overview

Journal Arch Microbiol

Specialty Microbiology

Date 2019 Aug 3

PMID 31372664

Citations 7

Authors

Mark Arentshorst

Davina de Lange

Joohae Park

Ellen L Lagendijk

Ebru Alazi

Cees A M J J Van Den Hondel

Arthur F J Ram

Affiliations

Soon will be listed here.

Abstract

Galactofuranose (Galf)-containing glycostructures are important to secure the integrity of the fungal cell wall. Golgi-localized Galf-transferases (Gfs) have been identified in Aspergillus nidulans and Aspergillus fumigatus. BLASTp searches identified three putative Galf-transferases in Aspergillus niger. Phylogenetic analysis showed that they group in three distinct groups. Characterization of the three Galf-transferases in A. niger by constructing single, double, and triple mutants revealed that gfsA is most important for Galf biosynthesis. The growth phenotypes of the ΔgfsA mutant are less severe than that of the ΔgfsAC mutant, indicating that GfsA and GfsC have redundant functions. Deletion of gfsB did not result in any growth defect and combining ΔgfsB with other deletion mutants did not exacerbate the growth phenotype. RT-qPCR experiments showed that induction of the agsA gene was higher in the ΔgfsAC and ΔgfsABC compared to the single mutants, indicating a severe cell wall stress response after multiple gfs gene deletions.

Citing Articles

Substrate binding and catalytic mechanism of UDP-α-D-galactofuranose: β-galactofuranoside β-(1→5)-galactofuranosyltransferase GfsA.

Oka T, Okuno A, Hira D, Teramoto T, Chihara Y, Hirata R PNAS Nexus. 2024; 3(11):pgae482.

PMID: 39507050 PMC: 11538602. DOI: 10.1093/pnasnexus/pgae482.

Identification of galactofuranose antigens such as galactomannoproteins and fungal-type galactomannan from the yellow fungus ().

Kadooka C, Tanaka Y, Hira D, Maruyama J, Goto M, Oka T Front Microbiol. 2023; 14:1110996.

PMID: 36814571 PMC: 9939772. DOI: 10.3389/fmicb.2023.1110996.

Galactofuranose (Galf)-containing sugar chain contributes to the hyphal growth, conidiation and virulence of F. oxysporum f.sp. cucumerinum.

Zhou H, Xu Y, Ebel F, Jin C PLoS One. 2021; 16(7):e0250064.

PMID: 34329342 PMC: 8323920. DOI: 10.1371/journal.pone.0250064.

Current Practices for Reference Gene Selection in RT-qPCR of Aspergillus: Outlook and Recommendations for the Future.

Archer M, Xu J Genes (Basel). 2021; 12(7).

PMID: 34202507 PMC: 8307107. DOI: 10.3390/genes12070960.

Galactomannan Produced by : An Update on the Structure, Biosynthesis and Biological Functions of an Emblematic Fungal Biomarker.

Fontaine T, Latge J J Fungi (Basel). 2020; 6(4).

PMID: 33198419 PMC: 7712326. DOI: 10.3390/jof6040283.

References

Punt P, van den Hondel C . Transformation of filamentous fungi based on hygromycin B and phleomycin resistance markers. Methods Enzymol. 1992; 216:447-57. DOI: 10.1016/0076-6879(92)16041-h. View

Heesemann L, Kotz A, Echtenacher B, Broniszewska M, Routier F, Hoffmann P . Studies on galactofuranose-containing glycostructures of the pathogenic mold Aspergillus fumigatus. Int J Med Microbiol. 2011; 301(6):523-30. DOI: 10.1016/j.ijmm.2011.02.003. View

El-Ganiny A, Sanders D, Kaminskyj S . Aspergillus nidulans UDP-galactopyranose mutase, encoded by ugmA plays key roles in colony growth, hyphal morphogenesis, and conidiation. Fungal Genet Biol. 2008; 45(12):1533-42. DOI: 10.1016/j.fgb.2008.09.008. View

El-Ganiny A, Sheoran I, Sanders D, Kaminskyj S . Aspergillus nidulans UDP-glucose-4-epimerase UgeA has multiple roles in wall architecture, hyphal morphogenesis, and asexual development. Fungal Genet Biol. 2010; 47(7):629-35. DOI: 10.1016/j.fgb.2010.03.002. View

Engel J, Schmalhorst P, Dork-Bousset T, Ferrieres V, Routier F . A single UDP-galactofuranose transporter is required for galactofuranosylation in Aspergillus fumigatus. J Biol Chem. 2009; 284(49):33859-68. PMC: 2797156. DOI: 10.1074/jbc.M109.070219. View

Park J, Hulsman M, Arentshorst M, Breeman M, Alazi E, Lagendijk E . Transcriptomic and molecular genetic analysis of the cell wall salvage response of Aspergillus niger to the absence of galactofuranose synthesis. Cell Microbiol. 2016; 18(9):1268-84. PMC: 5129474. DOI: 10.1111/cmi.12624. View

Broekhuijsen M, Unkles S, CAMPBELL E, Kinghorn J, Contreras R, Pouwels P . A gene transfer system based on the homologous pyrG gene and efficient expression of bacterial genes in Aspergillus oryzae. Curr Genet. 1989; 16(3):159-63. DOI: 10.1007/BF00391472. View

Lee M, Gravelat F, Cerone R, Baptista S, Campoli P, Choe S . Overlapping and distinct roles of Aspergillus fumigatus UDP-glucose 4-epimerases in galactose metabolism and the synthesis of galactose-containing cell wall polysaccharides. J Biol Chem. 2013; 289(3):1243-56. PMC: 3894311. DOI: 10.1074/jbc.M113.522516. View

Toledo M, Levery S, Bennion B, Guimaraes L, Castle S, Lindsey R . Analysis of glycosylinositol phosphorylceramides expressed by the opportunistic mycopathogen Aspergillus fumigatus. J Lipid Res. 2007; 48(8):1801-24. DOI: 10.1194/jlr.M700149-JLR200. View

10.

Park J, Tefsen B, Arentshorst M, Lagendijk E, van den Hondel C, Van Die I . Identification of the UDP-glucose-4-epimerase required for galactofuranose biosynthesis and galactose metabolism in . Fungal Biol Biotechnol. 2017; 1:6. PMC: 5598270. DOI: 10.1186/s40694-014-0006-7. View

11.

Arentshorst M, Ram A, Meyer V . Using non-homologous end-joining-deficient strains for functional gene analyses in filamentous fungi. Methods Mol Biol. 2011; 835:133-50. DOI: 10.1007/978-1-61779-501-5_9. View

12.

Schmalhorst P, Krappmann S, Vervecken W, Rohde M, Muller M, Braus G . Contribution of galactofuranose to the virulence of the opportunistic pathogen Aspergillus fumigatus. Eukaryot Cell. 2008; 7(8):1268-77. PMC: 2519766. DOI: 10.1128/EC.00109-08. View

13.

Damveld R, Franken A, Arentshorst M, Punt P, Klis F, Van Den Hondel C . A novel screening method for cell wall mutants in Aspergillus niger identifies UDP-galactopyranose mutase as an important protein in fungal cell wall biosynthesis. Genetics. 2008; 178(2):873-81. PMC: 2248356. DOI: 10.1534/genetics.107.073148. View

14.

Afroz S, El-Ganiny A, Sanders D, Kaminskyj S . Roles of the Aspergillus nidulans UDP-galactofuranose transporter, UgtA in hyphal morphogenesis, cell wall architecture, conidiation, and drug sensitivity. Fungal Genet Biol. 2011; 48(9):896-903. DOI: 10.1016/j.fgb.2011.06.001. View

15.

Bardalaye P, Nordin J . Chemical structure of the galactomannan from the cell wall of Aspergillus niger. J Biol Chem. 1977; 252(8):2584-91. View

16.

Wallis G, Swift R, Hemming F, Trinci A, Peberdy J . Glucoamylase overexpression and secretion in Aspergillus niger: analysis of glycosylation. Biochim Biophys Acta. 1999; 1472(3):576-86. DOI: 10.1016/s0304-4165(99)00188-9. View

17.

Park J, Tefsen B, Heemskerk M, Lagendijk E, Van Den Hondel C, Van Die I . Identification and functional analysis of two Golgi-localized UDP-galactofuranose transporters with overlapping functions in Aspergillus niger. BMC Microbiol. 2015; 15:253. PMC: 4630932. DOI: 10.1186/s12866-015-0541-2. View

18.

Oka T . Biosynthesis of galactomannans found in filamentous fungi belonging to Pezizomycotina. Biosci Biotechnol Biochem. 2018; 82(2):183-191. DOI: 10.1080/09168451.2017.1422383. View

19.

Lamarre C, Beau R, Balloy V, Fontaine T, Wong Sak Hoi J, Guadagnini S . Galactofuranose attenuates cellular adhesion of Aspergillus fumigatus. Cell Microbiol. 2009; 11(11):1612-23. DOI: 10.1111/j.1462-5822.2009.01352.x. View

20.

Katafuchi Y, Li Q, Tanaka Y, Shinozuka S, Kawamitsu Y, Izumi M . GfsA is a β1,5-galactofuranosyltransferase involved in the biosynthesis of the galactofuran side chain of fungal-type galactomannan in Aspergillus fumigatus. Glycobiology. 2017; 27(6):568-581. DOI: 10.1093/glycob/cwx028. View