title: Cold-induced cysts of the photosynthetic dinoflagellate Lingulodinium polyedrum have an arrested circadian bioluminescence rhythm and lower levels of protein phosphorylation.
abstract: Dinoflagellates are microscopic, eukaryotic, and primarily marine plankton. Temporary cyst formation is a well-known physiological response of dinoflagellate cells to environmental stresses. However, the molecular underpinnings of cold-induced cyst physiology have never been described. Cultures of the photosynthetic dinoflagellate Lingulodinium polyedrum readily form temporary cysts when placed at low (8°C±1°C) temperature and excyst to form normal motile cells following a return to normal temperature (18°C±1°C). The normal circadian bioluminescence rhythm and the expected changes in Luciferin Binding Protein abundance were arrested in L. polyedrum cysts. Furthermore, after excystment, the bioluminescence rhythm initiates at a time corresponding to zeitgeber 12, independent of the time when the cells encysted. Phosphoprotein staining after two-dimensional polyacrylamide gel electrophoresis, as well as column-based phosphoprotein enrichment followed by liquid chromatography tandem mass spectrometry, showed cyst proteins are hypophosphorylated when compared with those from motile cells, with the most marked decreases found for predicted Casein Kinase2 target sites. In contrast to the phosphoproteome, the cyst proteome is not markedly different from motile cells, as assessed by two-dimensional polyacrylamide gel electrophoresis. In addition to changes in the phosphoproteome, RNA sequencing revealed that cysts show a significant decrease in the levels of 132 RNAs. Of the 42 RNAs that were identified by sequence analysis, 21 correspond to plastid-encoded gene products and 11 to nuclear-encoded cell wall/plasma membrane components. Our data are consistent with a model in which the highly reduced metabolism in cysts is achieved primarily by alterations in the phosphoproteome. The stalling of the circadian rhythm suggests temporary cysts may provide an interesting model to address the circadian system of dinoflagellates.
authors: Département de Sciences Biologiques, Institut de Recherche en Biologie Végétale, Université de Montréal, 4101 Sherbrooke est, Montreal, Quebec, Canada H1X 2B2.
authors: Sougata
authors: S
authors: Roy
authors: Louis
authors: L
authors: Letourneau
authors: David
authors: D
authors: Morse
mesh_terms: D001683:Biological Clocks; D047390:Casein Kinase II / Q000378:metabolism; D002465:Cell Movement; D002940:Circadian Rhythm; D003080:Cold Temperature; D004141:Dinoflagellida / Q000502:physiology* / Q000648:ultrastructure; D015536:Down-Regulation / Q000235:genetics; D049449:Luminescence; D010748:Phosphopeptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010788:Photosynthesis; D018087:Plastids / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA
chemical_list: D010748:Phosphopeptides; D010750:Phosphoproteins; D015800:Protozoan Proteins; D018749:RNA, Plant; D047390:Casein Kinase II