Characterization of cell-cycle-driven and light-driven gene expression in a synchronous culture system in the unicellular rhodophyte Cyanidioschyzon merolae.
The unicellular rhodophyte Cyanidioschyzon merolae, having a single plastid and a single mitochondrion, is suitable for the analysis of the cell cycle involving the division of organelles. In conventional methods of synchronous culture of algae, light/dark cycles have been used as signals for synchronization, and the gene expression promoted by light is not separated from the gene expression related to cell cycle progression. We previously devised a novel synchronous culture system with controlled photosynthesis, which is triggered by 6 h-light/18 h-dark cycles combined with different levels of CO(2). The cells do not enter S-phase and consequently do not divide after the minimum light period without CO(2) supplementation, but do divide after a light period with 1 % CO(2). In this way, we can compare a dividing cycle and a non-dividing cycle. We examined changes in the expression of 74 genes throughout the cell cycle by quantitative RT-PCR. The expression of genes for two cyclins (cyclin C and H) and two CDKs (CDKA and CDKD) as well as metabolic enzymes was promoted by light, whereas the expression of genes for G1/S or G2/M cyclins and CDKs as well as DNA replication enzymes and proteins related to organellar division was promoted only in the dividing cycles. These results suggested that C. merolae has a checkpoint for G1/S progression, which is regulated by nutrients within the 6 h light period.
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.|Department of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
Takashi
T
Moriyama
Graduate School of Life Sciences, Tohoku University, Katahira, Aoba-ku, Sendai 980-8577, Japan.|Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Kimihiro
K
Terasawa
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Kohsuke
K
Sekine
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Masakazu
M
Toyoshima
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Mika
M
Koike
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Makoto
M
Fujiwara
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
Naoki
N
Sato
D002453:Cell Cycle; D046508:Culture Techniques; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism; D016213:Cyclins / Q000235:genetics / Q000378:metabolism; D048028:DNA, Algal / Q000032:analysis; D020869:Gene Expression Profiling; D008027:Light; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D010788:Photosynthesis; D018087:Plastids / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000461:Rhodophyta / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism; D016196:S Phase
D016213:Cyclins; D048028:DNA, Algal; D018844:Cyclin-Dependent Kinases