Shi Xinguo, Ma Minglei, Lin Senjie
State Key Laboratory of Marine Environmental Science, College of Ocean and Earth Sciences, Xiamen UniversityXiamen, China.
Department of Marine Sciences, University of Connecticut, GrotonCT, United States.
Front Microbiol. 2017 Jun 20;8:1118. doi: 10.3389/fmicb.2017.01118. eCollection 2017.
Dinoflagellates undergo a typical eukaryotic cell cycle consisting of G1, S, G2, and M phases and some of the typical cell cycle related genes have been computationally identified. However, very few of these genes have been experimentally linked to the cell cycle phases. Besides, although thecate dinoflagellates are known to possess theca composed of cellulose, information on cellulose synthesis and degradation associated with the cell cycle is also limited. In this study, we isolated G1/S cyclin, cellulose synthase and cellulase encoding genes in dinoflagellate . Further, using reverse transcription quantitative PCR (RT-qPCR), we characterized the expression profiles of the three genes throughout the cell cycle. All three showed clear expression dynamics throughout the cell cycle, with fold changes of 26, 2.4 and 9.3 for G1/S cyclin, cellulose synthase and cellulase gene, respectively. The transcript abundance of G1/S cyclin increased in late G1 phase and dropped in early S phase, indicating that this protein is involved in the G1/S transition. Throughout the cell cycle, the average transcript level of cellulose synthase was 4.5-fold higher than that of cellulase. Cellulose synthase and cellulase gene expressions showed peak transcript abundances at middle G1 phase and G2M phase, respectively, indicating the respective roles of these enzymes in the growth of newly divided cells and in cytokinesis. Our results suggest that G1/S cyclin, cellulase, and cellulose synthase genes associated with G1/S transition, G2M, and G1 phases of the cell cycle and are candidates of biomarkers for assessing growth status of .
甲藻经历典型的真核细胞周期，包括G1、S、G2和M期，并且一些典型的与细胞周期相关的基因已通过计算鉴定出来。然而，这些基因中很少有在实验上与细胞周期阶段相关联。此外，尽管已知有壳甲藻具有由纤维素组成的壳，但与细胞周期相关的纤维素合成和降解的信息也很有限。在本研究中，我们在甲藻中分离出了编码G1/S细胞周期蛋白、纤维素合酶和纤维素酶的基因。此外，我们使用逆转录定量PCR（RT-qPCR）对这三个基因在整个细胞周期中的表达谱进行了表征。所有这三个基因在整个细胞周期中均显示出明显的表达动态，G1/S细胞周期蛋白、纤维素合酶和纤维素酶基因的倍数变化分别为26、2.4和9.3。G1/S细胞周期蛋白的转录本丰度在G1期后期增加，在S期早期下降，表明该蛋白参与G1/S转变。在整个细胞周期中，纤维素合酶的平均转录水平比纤维素酶高4.5倍。纤维素合酶和纤维素酶基因的表达分别在G1期中期和G2M期显示出转录本丰度峰值，表明这些酶在新分裂细胞的生长和胞质分裂中的各自作用。我们的结果表明，与细胞周期的G1/S转变、G2M和G1期相关的G1/S细胞周期蛋白、纤维素酶和纤维素合酶基因，是评估甲藻生长状态的生物标志物候选基因。
