AG Hippler - (vi) N-glycosylation in algae

The N-glycosylation pathway in C. reinhardtii was disentangled in 2013 and many enzymes have been suggested as putative N-glycosyltransferases (Mathieu-Rivet et al., 2013). In recent work we revealed the function of two candidate genes (Man1A and XylT1A) in N-glycosylation of C. reinhardtii (Schulze et al., 2018) and analyzed the role of two other putative glycosyltransferases (FucT encoded as Cre18.g749697 and XylT1B encoded as Cre16.g678997) (Lucas et al., 2020, Oltmanns et al., 2020). Taking advantage of single-cell adhesion force measurements, our data revealed that cell adhesion was significantly impaired in C. reinhardtii N-glycosylation mutant strains (Man1A, XylT1A and a double mutant generated by genetic crossing) as compared to WT (Xu et al., 2020). Our data further suggested that flagellar assembly (data not shown), intraflagellar transport (IFT) and FMG1-B transport into flagella were not affected by altered N-glycosylation. Thus, we suggest that N-glycosylation of flagellar proteins is crucial for adhering C. reinhardtii cells onto surfaces. To further elucidate this phenotype, we aim to continue our collaborative adhesion force measurements using mutant strains which are devoid of core xylose and fucose (Oltmanns et al., 2020).

Xu N, Oltmanns A, Zhao L, Girot A, Karimi M, Hoepfner L, Kelterborn S, Scholz M, Beißel J, Hegemann P, Bäumchen O, Liu LN, Huang K, Hippler M (2020)
Altered N-glycan composition impacts flagella mediated adhesion in Chlamydomonas reinhardti.
Elife in press

Schulze S, Oltmanns A, Fufezan C, Krägenbring J, Mormann M, Pohlschroder M, Hippler M (2020)
SugarPy facilitates the universal, discovery-driven analysis of intact glycopeptides.
Bioinformatics in press

Oltmanns A, Hoepfner L, Scholz M, Zinzius K, Schulze S, Hippler M (2020)
Novel Insights Into N-Glycan Fucosylation and Core Xylosylation in C. reinhardtii.
Front Plant Sci 10: 1686

Lucas PL, Mathieu-Rivet E, Song PCT, Oltmanns A, Loutelier-Bourhis C, Plasson C, Afonso C, Hippler M, Lerouge P, Mati-Baouche N, Bardor M (2020)
Multiple xylosyltransferases heterogeneously xylosylate protein N-linked glycans in Chlamydomonas reinhardtii.
Plant J 102: 230-245

Schulze S, Oltmanns A, Machnik N, Liu G, Xu N, Jarmatz N, Scholz M, Sugimoto K, Fufezan C, Huang K, Hippler M (2018)
N-Glycoproteomic Characterization of Mannosidase and Xylosyltransferase Mutant Strains of Chlamydomonasreinhardtii.
Plant Physiol 176: 1952-1964

Schulze S, Urzica E, Reijnders M, van de Geest H, Warris S, Bakker LV, Fufezan C, Martins Dos Santos VAP, Schaap PJ, Peters SA, Hippler M (2017)
Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni.
New Phytol 215: 1361-1369

Mathieu-Rivet E, Scholz M, Arias C, Dardelle F, Schulze S, Le Mauff F, Teo G, Hochmal AK, Blanco-Rivero A, Loutelier-Bourhis C, Kiefer-Meyer MC, Fufezan C, Burel C, Lerouge P, Martinez F, Bardor M, Hippler M (2013)
Exploring the N-glycosylation pathway in Chlamydomonas reinhardtii unravels novel complex structures.
Molecular & cellular proteomics : MCP 12: 3160-83