LUO Qiulan,WANG Chaogang,and HU Zhangli.Bioinformatical analysis of genes encoding APC/C complex and the expression profiles analysis of CrCDC20 in Chlamydomanos reinhardtii[J].Journal of Shenzhen University Science and Engineering,2018,35(2):172-178.[doi:10.3724/SP.J.1249.2018.02172]





Bioinformatical analysis of genes encoding APC/C complex and the expression profiles analysis of CrCDC20 in Chlamydomanos reinhardtii
1) 深圳大学生命与海洋科学学院,深圳市海洋生物资源与生态环境重点实验室,广东深圳 518060
2) 深圳大学光电工程学院,光电子器件与系统教育部/广东省重点实验室,广东深圳 518060
3) 深圳大学龙华生物产业创新研究院,深圳市海洋藻类生物工程技术研究中心,广东深圳 518060
4) 中国热带农业科学院热带生物技术研究所,海南海口 570100
LUO Qiulan1234 WANG Chaogang13 and HU Zhangli13
1) College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Marine Bioresource & Eco-environmental Sciences, Shenzhen University, henzhen 518060, Guangdong Province, P.R.China
2) College of Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen, Guangdong 518060, Guangdong Province, P.R.China
3) Shenzhen Engineering Laboratory for Marine Algal Biotechnology, Longhua Innovation Institute for Biotechnology, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R China
4) Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Science, Haikou 570100, Hainan Province, P.R.China
molecular biology of microalgae biodiesel cell cycle E3 ubiquitin ligase phylogenetic analysis real-time fluorescence quantitative reverse transcription polymerase chain reaction (rt-RT-qPCR)
Q 946.1
为揭示莱茵衣藻细胞周期调控机制,提高微藻生物量,通过生物信息学方法,对莱茵衣藻参与细胞周期控制的末期促进复合物(anaphase-promoting complex/cyclosome,APC/C)基因家族的性质进行系统分析,包括基因结构、蛋白质特性和进化树分析等. 结果显示,莱茵衣藻包含13个APC/C复合物编码基因,在13号染色体存在一个基因簇. 除了激活因子细胞分裂周期蛋白20(cell division cycle 20,CDC20)和CDC20同系蛋白1(CDC20 homolog 1,CDH1)相对保守外,莱茵衣藻APC/C复合物的其他亚基与高等植物来源的蛋白质保守性很低. 此外,利用实时荧光定量反转录聚合酶链式反应(real-time fluorescence quantitative reverse transcription polymerase chain reaction, rt-RT-qPCR)分析了莱茵衣藻CrCDC20基因在缺氮或缺硫下表达谱,发现CrCDC20基因在缺硫下上调表达,在缺氮下下调表达,说明CrCDC20参与莱茵衣藻的营养缺乏胁迫应答反应.
Chlamydomanos reinhardtii is a model microalgae for lipids production. There is a contradiction between lipids biosynthesis and biomass accumulation in C. reinhardtii. Revealing regulation mechanisms of cell cycle can prove a new direction for promoting biomass in C. reinhardtii. In this paper, the gene structures, protein characteristics, and phylogenetic tree of anaphase-promoting complex/cyclosome (APC/C) gene family involving in cell cycle control are analyzed through bioinformatics. Totally 13 genes of APC/C family are identified in C. reinhardtii, and one gene cluster is found in chromosome 13. The phylogenetic tree shows that APC/C proteios, except the activation factors CrCDC20 and CrCDH1 from C. reinhardtii, have low homologous with the proteios in higher plants. Meanwhile, the gene expression profiles of CrCDC20 under sulfur or nitrogen deficiency conditions are determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (rt-RT-qPCR). The results show that CrCDC20 is up-regulate under sulfur deficiency but down-regulate under nitrogen deficiency, which indicates that CrCDC20 might involve in nutrition deficiency stresses.


[1] RATHA S K, PRASANNA R. Bioprospecting microalgae as potential sources of “green energy”—challenges and perspectives (review)[J]. Applied Biochemistry & Microbiology, 2012, 48(2):109-125.
[2] DENG Xiaodong, FAN Xinzhao, LI Ping, et al. A photoperiod-regulating gene CONSTANS is correlated to lipid biosynthesis in Chlamydomonas reinhardtii[J]. BioMed Research International, 2015(4): 715020.
[3] GHASEMI Y, RASOULAMINI S, NASERI A T, et al. Microalgae biofuel potentials (review)[J]. Applied Biochemistry & Microbiology, 2012, 48(2):150-168.
[4] MOLNAR A, BASSETT A, THUENEMANN E, et al. Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii[J]. Plant Cell, 2006, 58(5): 1121-1133.
[5] YAMANO T, IGUCHI H, FUKUZAWA H. Rapid transformation of Chlamydomonas reinhardtii without cell-wall removal[J]. Journal of Bioscience & Bioengineering, 2013, 115(6): 691-694.
[6] L Hexin, QU Ge, QI Xizhen, et al. Transcriptome analysis of Chlamydomonas reinhardtii during the process of lipid accumulation[J]. Genomics, 2013, 101(4): 229-237.
[7] MILLER R, WU Guangxi, DESHPANDE R R, et al. Changes in transcript abundance in Chlamydomonas reinhardtii following nitrogen deprivation predict diversion of metabolism[J]. Plant Physiology, 2010, 154(4): 1737-1752.
[8] OUYANG Longling, LI Hui, YAN Xiaojun, et al. Site-directed mutagenesis from Arg195 to His of a microalgal putatively chloroplastidial glycerol-3-phosphate acyltransferase causes an increase in phospholipid levels in yeast[J]. Frontiers in Plant Science, 2016: 7. doi: https://doi.org/10.3389/fpls.2016.00286.
[9] YOON K, HAN Danxiang, LI Yantao, et al. Phospholipid:diacylglycerol acyltransferase is a multifunctional enzyme involved in membrane lipid turnover and degradation while synthesizing triacylglycerol in the unicellular green microalga Chlamydomonas reinhardtii[J]. Plant Cell, 2012, 24(9): 3708-3724.
[10] TAN K W, LEE Y K. Expression of the heterologous Dunaliella tertiolecta fatty acyl-ACP thioesterase leads to increased lipid production in Chlamydomonas reinhardtii[J]. Journal of Biotechnology, 2017, 247:60-67.
[11] DRIVER T, TRIVEDI D K, MCINTOSH O A, et al. Two glycerol-3-phosphate dehydrogenases from Chlamydomonas have distinct roles in lipid metabolism[J]. Plant Physiology, 2017, 174(4):2083-2097.
[12] IWAI M, IKEDA K, SHIMOJIMA M, et al. Enhancement of extraplastidic oil synthesis in Chlamydomonas reinhardtii using a type-2 diacylglycerol acyltransferase with a phosphorus starvation-inducible promoter[J]. Plant Biotechnology Journal, 2014, 12(6): 808-819.
[13] KAO P H, NG I S. CRISPRi mediated phosphoenolpyruvate carboxylase regulation to enhance the production of lipid in Chlamydomonas reinhardtii[J]. Bioresource Technology, 2017, 245(Pt B):1527-1537.
[14] 翟中和, 王喜忠, 丁明孝. 细胞生物学[M].4版.北京:高等教育出版社, 2011: 358-371.
ZHAI Zhonghe, WANG Xizhong, DING Mingxiao. Cell Biology[M]. 4th edition. Beijing: Higher Education Press, 2011: 358-371.(in Chinese)
[15] ACQUAVIVA C, HERZOG F, KRAFT C, et al. The anaphase promoting complex/cyclosome is recruited to centromeres by the spindle assembly checkpoint[J]. Nature Cell Biology, 2004, 6(9):892-898.
[16] TANG Z Y, LI B, BHARADWAJ R, et al. APC2 Cullin protein and APC11 RING protein comprise the minimal ubiquitin ligase module of the anaphase-promoting complex[J]. Molecular Biology of the Cell, 2001, 12(12):3839-3851.
[17] VISINTIN R, PRINZ S, AMON A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis[J]. Science, 1997, 278(5337): 460-463.
[18] MORRIS M C, KAISER P, RUDYAK S, et al. Cks1-dependent proteasome recruitment and activation of CDC20 transcription in budding yeast[J]. Nature 2003, 423: 1009-1013.
[19] BIOV K, ZACHLEDER V. Cell-cycle regulation in green algae dividing by multiple fission[J]. Journal of Experimental Botany, 2014, 65(10): 2585-2602.
[20] CROSS F R, UMEN J G. The Chlamydomonas cell cycle[J]. Plant Journal, 2015, 82(3):370-392.
[21] JPPNER J, MUBEEN U, LEISSE A, et al. Dynamics of lipids and metabolites during the cell cycle of Chlamydomonas reinhardtii[J]. Plant Journal for Cell & Molecular Biology, 2017, 92: 331-343.
[22] DENG Xiaodong, FEI Xiaowen, LI Yajun. The effects of nutritional restriction on neutral lipid accumulation in Chlamydomonas and Chlorella[J]. African Journal of Microbiology Research, 2011, 5(3):260-270.
[23] CAKMAK T, ANGUN P, OZKAN A D, et al. Nitrogen and sulfur deprivation differentiate lipid accumulation targets of Chlamydomonas reinhardtii[J]. Bioengineered Bugs, 2012, 3(6):343-346.
[24] MARROCCO K, CRIQUI M C, ZERVUDACKI J, et al. APC/C-mediated degradation of dsRNA-binding protein 4 (DRB4) involved in RNA silencing[J]. PLOS One, 2012, 7(4):e35173.
[25] HEYMAN J, VEYLDER L D. The anaphase-promoting complex/cyclosome in control of plant development[J]. Molecular Plant, 2012, 5(6): 1182-1194.
[26] LEMOINE Y, SCHOEFS B. Secondary ketocarotenoid astaxanthin biosynthesis in algae: a multifunctional response to stress[J]. Photosynthesis Research, 2010, 106(1/2):155-177.


Foundation:National Natural Science Foundation of China (31700309, 31470389, 31470431); Natural Science Foundation of Guangdong Province (2014A030308017, 2017A030310255); Shenzhen Science and Technology Research Foundation (JCYJ20150324141711583)
Corresponding author:Professor HU Zhangli.E-mail: huzl@szu.edu.cn
Citation:LUO Qiulan,WANG Chaogang,HU Zhangli.Bioinformatical analysis of genes encoding APC/C complex and the expression profiles analysis of CrCDC20 in Chlamydomanos reinhardtii[J]. Journal of Shenzhen University Science and Engineering, 2018, 35(2): 172-178.(in Chinese)
基金项目:国家自然科学基金资助项目(31700309, 31470389, 31470431);广东省自然科学基金资助项目(2014A030308017, 2017A030310255);深圳市科技计划资助项目(JCYJ20150324141711583)
作者简介:罗秋兰(1984— ),女,深圳大学博士后研究人员,中国热带农业科学院助理研究员.研究方向:藻类分子生物学.E-mail:luoqiulan_79@szu.edu.cn
引文:罗秋兰,王潮岗,胡章立.莱茵衣藻APC/C复合物基因家族及CrCDC20表达谱分析[J]. 深圳大学学报理工版,2018,35(2):172-178.
更新日期/Last Update: 2018-03-07