The structural challenges faced by eukaryotic cells through the cell cycle are key for understanding cell viability and proliferation. We tested the hypothesis that the biosynthesis of structural lipids is linked to the cell cycle. If true, this would suggest that the cell's structure is important for progress through and perhaps even control of the cell cycle. Lipidomics (31P NMR and MS), proteomics (Western immunoblotting) and transcriptomics (RT-qPCR) techniques were used to profile the lipid fraction and characterise aspects of its metabolism at seven stages of the cell cycle of the model eukaryote, Desmodesmus quadricauda. We found considerable, transient increases in the abundance of phosphatidylethanolamine during the G1 phase (+35%, ethanolamine phosphate cytidylyltransferase increased 2·5×) and phosphatidylglycerol (+100%, phosphatidylglycerol synthase increased 22×) over the G1/pre-replication phase boundary. The relative abundance of phosphatidylcholine fell by ~35% during the G1. N-Methyl transferases for the conversion of phosphatidylethanolamine into phosphatidylcholine were not found in the de novo transcriptome profile, though a choline phosphate transferase was found, suggesting that the Kennedy pathway is the principal route for the synthesis of PC. The fatty acid profiles of the four most abundant lipids suggested that these lipids were not generally converted between one another. This study shows for the first time that there are considerable changes in the biosynthesis of the three most abundant phospholipid classes in the normal cell cycle of D. quadricauda, by margins large enough to elicit changes to the physical properties of membranes.

Department of Chemistry Universitetet i Oslo P O Box 1033 Blindern NO 0315 Oslo Norway

Department of Molecular Biology University of Bergen Thormøhlens gate 55 NO 5008 Bergen Norway

Department of Molecular Biology University of Bergen Thormøhlens gate 55 NO 5008 Bergen Norway; Core Metabolomics and Lipidomics Laboratory Wellcome Trust MRL Institute of Metabolic Science University of Cambridge Level 4 Pathology Building Addenbrooke's Hospital Cambridge CB2 0QQ United Kingdom; Biological chemistry group Jodrell laboratory Royal Botanic Gardens Kew United Kingdom

Laboratory of Cell Cycles of Algae Centre Algatech Institute of Microbiology of the Czech Academy of Sciences Novohradská 237 379 01 Třeboň Czech Republic

Laboratory of Cell Cycles of Algae Centre Algatech Institute of Microbiology of the Czech Academy of Sciences Novohradská 237 379 01 Třeboň Czech Republic; Department of Functional Ecology Institute of Botany of the Czech Academy of Sciences Dukelská 135 379 81 Třeboň Czech Republic

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