Most microalgae produce triacylglycerol (TAG) under stress conditions such as nitrogen depletion, but the underlying molecular mechanism remains unclear. In this study, we focused on the role of target of rapamycin (TOR) in TAG accumulation. TOR is a serine/threonine protein kinase that is highly conserved and plays pivotal roles in nitrogen and other signaling pathways in eukaryotes. We previously constructed a rapamycin-susceptible Cyanidioschyzon merolae, a unicellular red alga, by expressing yeast FKBP12 protein to evaluate the results of TOR inhibition (Imamura et al. in Biochem Biophys Res Commun 439:264-269, 2013). By using this strain, we here report that rapamycin-induced TOR inhibition results in accumulation of cytoplasmic lipid droplets containing TAG. Transcripts for TAG synthesis-related genes, such as glycerol-3-phosphate acyltransferase and acyl-CoA:diacylglycerol acyltransferase (DGAT), were increased by rapamycin treatment. We also found that fatty acid synthase-dependent de novo fatty acid synthesis was required for the accumulation of lipid droplets. Induction of TAG and up-regulation of DGAT gene expression by rapamycin were similarly observed in the unicellular green alga, Chlamydomonas reinhardtii. These results suggest the general involvement of TOR signaling in TAG accumulation in divergent microalgae.

No abstract

Cyanobacteria have developed a light-harvesting antenna complex known as the phycobilisome. When cells are starved for nutrients or exposed to high light, the phycobilisome is rapidly degraded (bleaching). It has been suggested that in the cyanobacterium Synechococcus elongatus PCC 7942, the bleaching process is regulated by a two-component histidine kinase, NblS. To clarify the signaling pathway involving NblS, we identified the NblS-interacting response regulators RpaB and SrrA. In vitro assays using recombinant proteins showed that both RpaB and SrrA can receive phosphoryl groups from autophosphorylated NblS; the NblS-interacting protein SipA clearly enhances the phosphotransfer activity from NblS to RpaB and SrrA. In addition, NblS prefers SrrA over RpaB as the phosphotransfer target with or without SipA. Gel mobility shift assay revealed that both RpaB and SrrA can bind to the upstream region of nblA, a major regulatory factor in the bleaching process. nblA transcript accumulates in nblS or rpaB mutants even under normal growth conditions, while in the srrA disruptant the nblA transcripts are slightly up-regulated under stress conditions. These observations suggest that the bleaching signal transduction pathway via NblS is regulated by RpaB and that SrrA is partially involved.

No abstract