高温胁迫下刺参H3K9ac特征及HATs、HDACs的表达  

The Acetylation Characteristics of H3K9 and the Expression of HATs and HDACs Under Heat Stress in Sea Cucumber(Apostichopus japonicus)

作　　者：张京京 徐冬雪 宋文琦 夏斌[2] 高勤峰[1] Zhang Jingjing;Xu Dongxue;Song Wenqi;Xia Bin;Gao Qinfeng(Key Laboratory of Mariculture(Ocean University of China),Ministry of Education,Qingdao 266003,China;Marine Science and Engineering College,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]海水养殖教育部重点实验室(中国海洋大学),山东青岛266003 [2]青岛农业大学海洋科学与工程学院,山东青岛266109

出　　处：《中国海洋大学学报（自然科学版）》2024年第2期44-54,共11页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31902360);山东省自然科学基金项目(ZR2022MC050)资助。

摘　　要：为了探索高温胁迫下刺参(Apostichopus japonicus)H3K9乙酰化水平的变化,研究组蛋白乙酰转移酶(Histone acetyltransferases,HATs)和去乙酰化酶(Histone deacetylases,HDACs)对刺参H3K9乙酰化的影响。本研究通过Western Blot技术研究高温胁迫条件下刺参肠道组织H3K9乙酰化特征,并对刺参基因组中4种组蛋白乙酰转移酶基因ajKAT2B、ajKAT5、ajKAT7、ajKAT8和4种组蛋白去乙酰化酶基因ajSIRT1、ajHDAC1、ajHDAC3、ajHDAC4进行结构解析和系统进化树分析;利用qRT-PCR定量研究在高温胁迫后刺参的8种基因表达量的变化模式。研究表明,通过Western Blot发现,在26℃胁迫下,H3K9ac水平在胁迫48 h后明显上升(P<0.05),在96 h后又迅速下降(P<0.05),表明其在高温胁迫下调控基因转录可能起到重要作用。基因结构解析发现HATs和HDACs含有保守的结构域,通过构建基因系统进化树发现其与物种进化树呈一致的趋势,也表现出功能上的高度保守。qRT-PCR结果表明:4种HATs基因在高温胁迫48 h后均有显著的上升(P<0.05),在胁迫96 h后,ajKAT2B和ajKAT8降低至对照组的表达水平,ajKAT5迅速下降至显著低于对照组水平,ajKAT7稍有降低但仍显著高于对照组。4种HDACs基因中,ajSIRT1在高温胁迫6 h后有明显的上升,在胁迫48 h后显著高于对照组(P<0.05),在胁迫96 h后降低至对照组的表达水平;ajHDAC1在高温胁迫6 h后有明显下降(P<0.05),在胁迫48 h后迅速上升至显著高于对照组水平(P<0.05),然后降低至对照组水平;ajHDAC3和ajHDAC4在高温胁迫6 h后均显著上升(P<0.05),在胁迫48 h后降至对照组水平并继续下降。研究结果表明,刺参H3K9ac乙酰化水平在高温胁迫下表现出明显动态变化,多种HATs和HDACs在mRNA水平上均有明显响应,这可能是导致H3K9ac变化的因素之一。The purpose of this study aims to explore the changes of acetylation levels of H3K9 in Apostiopsis japonicus under heat stress and the effects of HATs and HDACs on the changes.Western blot technique was used to determine the characteristics of H3K9ac at 26℃in A.japonicus.Through the analysis of the genome of A.japonicus,the gene structure and function domain of histone acetylase ajKAT2B,ajKAT5,ajKAT7,ajKAT8 and deacetylase ajSIRT1,ajHDAC1,ajHDAC3,ajHDAC4 were acquired.Phylogenetic trees were constructed by analyzing these 8 genes with those from other species,respectively.The temporal expression levels of the 8 genes in the intestine at 26℃were assessed by qRT-PCR.Western Blot analysis suggested that the abundance of H3K9ac gene transcript increased significantly after 48 h heat stressing,and decreased significantly after 96 h heat stressing(P<0.05),indicating that it may play an important role in regulating gene transcription under high temperature stress.The analysis of gene structure revealed that the evolution of the 8 genes was almost in accordance with the evolution of other species,which showed a high conservativeness in function.The results of qRT-PCR showed that after 48 h of high temperature stress,the mRNA abundances of the four HATs genes increased significantly(P<0.05),and after 96 h heat stress,the expression of ajKAT2B and ajKAT8 decreased to that of control.The expression of ajKAT5 decreased rapidly to the lowest,and the expression of ajKAT7 decreased slightly but was still significantly higher than that of control.Among the four HDACs,The mRNA abundance of ajSIRT1 increased significantly to the highest after 6 h heat stress,and still maintained a higher level than the control group after 48 h heat stress(P<0.05),and then decreased to the control level after 96 h heat stress.AjHDAC1 mRNA abundance decreased significantly after 6 h heat stress(P<0.05),and increased rapidly to the highest after 48 h heat stress(P<0.05),and then decreased to the level of the control group.The mRNA abundance of bo

关 键 词：刺参 高温胁迫 H3K9ac 组蛋白乙酰转移酶 组蛋白去乙酰化酶 

分 类 号：S968.9[农业科学—水产养殖]