Characterization of three non-canonical N-glycosylation motifs indicates N glyco -A reduces DNA N6-methyladenine and N glyco -D alters G/F actin ratio in Phytophthora sojae

Asparagine (Asn, N)-linked glycosylation is an abundant post-translational modification in which Asn, typically in N -X-S/T; X ≠ P motifs, are modified with N-glycans. It has essential regulatory roles in multicellular organisms. In this study, we systematically investigate the function of three N-g...

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Published in:International journal of biological macromolecules 2024-10, Vol.277 (Pt 2), p.133943
Main Authors: Chen, Shanshan, Wang, Yuke, Cui, Tongshan, Zheng, Yuxin, Zhang, Fan, Ma, Quanhe, Zhang, Can, Liu, Xili
Format: Article
Language:English
Subjects:
Actins - genetics
Actins - metabolism
Amino Acid Motifs
Autophagy
DNA Methylation
Glycosylation
Phytophthora - genetics
Phytophthora - metabolism
Phytophthora - pathogenicity
Protein Processing, Post-Translational
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Summary:Asparagine (Asn, N)-linked glycosylation is an abundant post-translational modification in which Asn, typically in N -X-S/T; X ≠ P motifs, are modified with N-glycans. It has essential regulatory roles in multicellular organisms. In this study, we systematically investigate the function of three N-glycosylation motifs (N -A, N -D and N -S) previously identified in Phytophthora sojae, through site-directed mutagenesis and functional assays. In P. sojae expressing glycosylation-dead variants pre-PsDMAP1 (N -A motif) or PsADF (N -D motif), zoospore release or cyst germination is impaired. In particular, the pre-PsDMAP1 mutant reduces DNA methylation levels, and the PsADF mutant disrupts the actin forms, which could explain the decrease in pathogenicity after N-glycosylation is destroyed. Similarly, P. sojae expressing PsNRX (N -S motif) shows increased sensitivity to H O and heat. Through autophagy or 26S proteasome pathway inhibition assays, we found that unglycosylated pre-PsDMAP1 and PsADF are degraded via the 26S proteasome pathway, while the autophagy pathway is responsible for PsNRX clearance. These findings demonstrate that glycosylation of these motifs regulates the stability and function of glycoproteins necessary for P. sojae growth, reproduction and pathogenicity, which expands the scope of known N-glycosylation regulatory functions in oomycetes.
ISSN:1879-0003