N-linked glycosylation enzymes in the diatom Thalassiosira oceanica exhibit a diel cycle in transcript abundance and favor for NXT-type sites
Open Access
- 5 February 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Scientific Reports
- Vol. 11 (1), 1-14
- https://doi.org/10.1038/s41598-021-82545-1
Abstract
N-linked glycosylation is a posttranslational modification affecting protein folding and function. The N-linked glycosylation pathway in algae is poorly characterized, and further knowledge is needed to understand the cell biology of algae and the evolution of N-linked glycosylation. This study investigated the N-linked glycosylation pathway in Thalassiosira oceanica, an open ocean diatom adapted to survive at growth-limiting iron concentrations. Here we identified and annotated the genes coding for the essential enzymes involved in the N-linked glycosylation pathway of T. oceanica. Transcript levels for genes coding for calreticulin, oligosaccharyltransferase (OST), N-acetylglucosaminyltransferase (GnT1), and UDP-glucose glucosyltransferase (UGGT) under high- and low-iron growth conditions revealed diel transcription patterns with a significant decrease of calreticulin and OST transcripts under iron-limitation. Solid-phase extraction of N-linked glycosylated peptides (SPEG) revealed 118 N-linked glycosylated peptides from cells grown in high- and low-iron growth conditions. The identified peptides had 81% NXT-type motifs, with X being any amino acids except proline. The presence of N-linked glycosylation sites in the iron starvation-induced protein 1a (ISIP1a) confirmed its predicted topology, contributing to the biochemical characterization of ISIP1 proteins. Analysis of extensive oceanic gene databases showed a global distribution of calreticulin, OST, and UGGT, reinforcing the importance of glycosylation in microalgae.Funding Information
- Deutscher Akademischer Austauschdienst
- Nova Scotia Graduate Stipend
- Natural Sciences and Engineering Research Council of Canada
- Discovery Grant
This publication has 69 references indexed in Scilit:
- N-Linked Glycosylation Is Required for Transferrin-Induced Stabilization of Transferrin Receptor 2, but Not for Transferrin Binding or Trafficking to the Cell SurfaceBiochemistry, 2013
- Green algae Chlamydomonas reinhardtii possess endogenous sialylated N‐glycansFEBS Open Bio, 2011
- Unique N-Glycan Moieties of the 66-kDa Cell Wall Glycoprotein from the Red Microalga Porphyridium sp.Online Journal of Public Health Informatics, 2011
- N-Glycans of Phaeodactylum tricornutum Diatom and Functional Characterization of Its N-Acetylglucosaminyltransferase I EnzymeOnline Journal of Public Health Informatics, 2011
- Mass spectrometric analysis of asparagine deamidation and aspartate isomerization in polypeptidesElectrophoresis, 2010
- Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotesMolecular Systems Biology, 2009
- Getting In and Out from Calnexin/Calreticulin CyclesOnline Journal of Public Health Informatics, 2008
- Solid-phase extraction of N-linked glycopeptidesNature Protocols, 2007
- Predicting transmembrane protein topology with a hidden markov model: application to complete genomesJournal of Molecular Biology, 2001
- KEGG: Kyoto Encyclopedia of Genes and GenomesNucleic Acids Research, 2000