林星桦,叶明慧,SEATAN Wanida,潘炎杨,陈芳圆,黄洋,朱春华,李广丽,田昌绪

• 1:广东海洋大学水产学院//广东省名特优鱼类生殖调控与繁育工程技术研究中心//南方海洋科学与工程广东省实验室(湛江)

摘要(Abstract)：

【目的】分析多鳞鱚(Sillago sihama)脯氨酸羟化酶(prolyl hydroxylases, Phds)基因家族序列特征,探讨其在低氧响应过程的作用。【方法】通过硬骨鱼类phds基因,在多鳞鱚转录组和基因组数据中比对筛选多鳞鱚phds基因。对多鳞鱚Phds蛋白序列进行结构域预测、系统进化和共线性分析等。采用实时荧光定量PCR检测phds基因在低氧胁迫和复氧后的表达变化。【结果】在多鳞鱚基因组中共鉴定到三个phds基因,分别是phd1、phd2和phd3,开放阅读框(ORF)全长分别为1 635、1 092、723 bp,分别编码544、363、240个氨基酸。结构域预测表明,多鳞鱚Phd1、Phd2、Phd3蛋白均含有P4Hc保守结构域。系统进化分析发现,多鳞鱚Phds均与大黄鱼(Larimichthys crocea)亲缘关系最近。共线性分析表明,多鳞鱚phds与尼罗罗非鱼(Oreochromis niloticus)上下游基因高度一致。phd1、phd2在低氧处理4 h鳃组织中m RNA表达显著高于常氧组(P <0.05),但在心脏中无显著性差异(P> 0.05);phd3在低氧处理4 h的鳃组织中显著低于常氧组(P <0.05),而在心脏中显著高于常氧组(P <0.05)。【结论】多鳞鱚存在phd1、phd2和phd3基因,并在不同组织的低氧响应中起重要作用。

关键词(KeyWords)： 多鳞鱚;phds基因;基因家族;低氧胁迫;基因表达

Abstract:

Keywords:

基金项目(Foundation): 广东省普通高校特色创新类项目(2019KTSCX060);; 广东省普通高校青年创新人才类项目(2018KQCX111);; 广东省科技计划项目(2014A020208117,2015A020209163);; 广东省大学生创新创业计划项目(CXXL2018048,CXXL2019138);; 广东海洋大学科研启动经费项目(R19026)

作者(Author): 林星桦,叶明慧,SEATAN Wanida,潘炎杨,陈芳圆,黄洋,朱春华,李广丽,田昌绪

参考文献(References)：

• [1] POLLOCK M S, CLARKE L M J, DUBé M G. The effects of hypoxia on fishes:from ecological relevance to physiological effects[J]. Environmental Reviews, 2007,15(NA):1-14.
• [2] ROBERTS J J, BRANDT S B, FANSLOW D, et al. Effects of hypoxia on consumption, growth, and RNA:DNA ratios of young yellow perch[J]. Transactions of the American Fisheries Society, 2011, 140(6):1574-1586.
• [3]肖武汉.低氧信号传导途径与鱼类低氧适应[J].中国科学:生命科学，2014,44(12):1227-1235.
• [4] DUNWOODIE S L. The role of hypoxia in development of the mammalian embryo[J]. Developmental Cell, 2009,17(6):755-773.
• [5]李福祥，全燕，夏前明.低氧诱导因子及其低氧调节机制[J].中国临床康复，2004,8(30):6744-6746.
• [6] SINGH L, ALDOSARY S, SAEEDAN A S, et al. Prolyl hydroxylase 2:a promising target to inhibit hypoxia-induced cellular metabolism in cancer cells[J].Drug Discovery Today, 2018, 23(11):1873-1882.
• [7] RYTK?NEN K T, WILLIAMS T A, RENSHAW G M, et al. Molecular evolution of the metazoan PHD-HIF oxygen-sensing system[J]. Molecular Biology and Evolution, 2011, 28(6):1913-1926.
• [8] LANDáZURI M O, VARA-VEGA A, VITóN M, et al.Analysis of HIF-prolyl hydroxylases binding to substrates[J]. Biochemical and Biophysical Research Communications, 2006, 351(2):313-320.
• [9] BERRA E, BENIZRI E, GINOUVèS A, et al. HIF prolyl-hydroxylase 2 is the key oxygen sensor setting low steady-state levels of HIF-1alpha in normoxia[J]. The EMBO Journal, 2003, 22(16):4082-4090.
• [10]陈楠.团头鲂phds基因家族参与低氧应答的分子机理解析[D].武汉:华中农业大学，2017.
• [11]薛泰强.鱚科几种鱼类的形态学及遗传学研究[D].青岛:中国海洋大学，2010.
• [12]黄洋，杜涛，黄海立.多鳞鱚Sillago sihama Forskál人工繁殖研究[J].广东海洋大学学报，2013,33(01):15-21.
• [13]杜涛，黄洋.多鳞鱚生物学特性及室内养殖试验[J].水产养殖，2009,30(3):1-3.
• [14]杜涛，黄洋，曹剑香.多鳞鱚采捕暂养的初步研究[J].养殖与饲料，2009(10):15-17.
• [15] LI Z Y, TIAN C X, HUANG Y, et al. A first insight into a draft genome of silver Sillago(Sillago sihama)via genome survey sequencing[J]. Animal, 2019, 9(10):756.
• [16] SAETAN W, TIAN C X, YU J W, et al. Comparative transcriptome analysis of gill tissue in response to hypoxia in silver Sillago(Sillago sihama)[J]. Animals,2020, 10(4):628.
• [17] ARAVIND L, KOONIN E V. The DNA-repair protein Alk B, EGL-9, and leprecan define new families of2-oxoglutarate-and iron-dependent dioxygenases[J].Genome Biology, 2001, 2(3):1-8.
• [18] GROSS C T, MCGINNIS W. DEAF-1, a novel protein that binds an essential region in a Deformed response element[J]. The EMBO Journal, 1996, 15(8):1961-1970.
• [19] HIRSIL?M, KOIVUNEN P, GüNZLER V, et al.Characterization of the human prolyl 4-hydroxylases that modify the hypoxia-inducible factor[J]. The Journal of Biological Chemistry, 2003, 278(33):30772-30780.
• [20] APPELHOFF R J, TIAN Y M, RAVAL R R, et al.Differential function of the prolyl hydroxylases PHD1,PHD2, and PHD3 in the regulation of hypoxia-inducible factor[J]. The Journal of Biological Chemistry, 2004,279(37):38458-38465.
• [21] LUOMA L M, BERRY F B. Molecular analysis of NPAS3 functional domains and variants[J]. BMC Molecular Biology, 2018, 19(1):1-19.
• [22] CHEN N, HUANG C X, HUANG C H, et al. The molecular characterization, expression pattern and alternative initiation of Megalobrama amblycephala Hif prolyl hydroxylase Phd1[J]. Gene, 2018, 678:219-225.
• [23] WANG H J, HUANG C X, CHEN N, et al. Molecular characterization and m RNA expression of HIF-prolyl hydroxylase-2(phd2)in hypoxia-sensing pathways from Megalobrama amblycephala[J]. Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 2015, 186:28-35.
• [24] CHEN N, HUANG C H, CHEN B X, et al. Alternative splicing transcription of Megalobrama amblycephala HIF prolyl hydroxylase PHD3 and up-regulation of PHD3 by HIF-1α[J]. Biochemical&Biophysical Research Communications, 2016, 469(3):737-742.