摘要
旨在克隆牦牛甲基转移酶样蛋白(methyltransferase-like protein 14,METTL14)基因的编码区(CDS)序列,探究其分子特征及时空表达规律。以大通牦牛脂肪组织cDNA为模板,通过克隆获得METTL14的完整CDS区序列,用生物学软件进行基因序列和蛋白质结构分析,利用qRT-PCR检测METTL14的时空表达规律。结果显示,大通牦牛METTL14的CDS区全长1371 bp,共编码456个氨基酸;METTL14亲缘关系与野牦牛最近,与鸡和小鼠最远;氨基酸等电点是5.89,不稳定系数(Ⅱ)为52.02,GRAVY=-0.878,METTL14总体表现出强的亲水性;METTL14有1个大小为177 aa的MT-A70酶保守结构域,无信号肽与跨膜结构域;亚细胞定位与核定位预测分析都显示METTL14位于细胞核(69.60%)。推测METTL14共有40个潜在的磷酸化位点。METTL14蛋白高级结构由无规则卷曲(51.10%)、α-螺旋(33.11%)、延伸链(11.84%)和β-转角(3.95%)相互连接而成。METTL14与甲基化修饰相关蛋白相互作用,主要参与RNA甲基化修饰过程。qRT-PCR结果显示,METTL14基因在牦牛7个组织中均有表达,相比脂肪组织中的表达,背最长肌中表达量最高(P<0.001);在牦牛生长发育的不同阶段,前体脂肪细胞中METTL14的表达量也有差异,30月龄>18月龄(P<0.05);进一步研究发现,在牦牛前体脂肪细胞分化过程中,METTL14表达量在0,4,8,12 d呈现上升的趋势,在4,8 d的表达量显著高于0 d(P<0.05),12 d的表达量极显著高于0 d(P<0.01)。结果表明,大通牦牛METTL14是一个具有MT-A70酶保守结构域的定位在细胞核上的酸性不稳定水溶性蛋白,在牦牛脂肪沉积过程中发挥着重要的作用。本试验进一步为METTL14在牦牛脂肪沉积中的调控机制研究奠定理论基础。
The aim of this study is to clone the sequence of the coding region(CDS)of yak methyltransferase-like protein 14(METTL14)and then to investigate its molecular characteristics and spatiotemporal expression regularity.The Datong yak adipose tissue cDNA was used as the template to clone the complete CDS sequence,and the METTL14 sequence and protein structure were analyzed by biological software,and the spatiotemporal expression pattern of METTL14 was detected by qRT-PCR.The results showed that the total length of CDS region of METTL14 is 1371 bp,encoding 456 amino acids;METTL14 is most closely related to wild yak and most distant to chicken and mouse.The amino acid isoelectric point of METTL14 is 5.89,and the instability coefficient(Ⅱ)is 52.02.METTL14 has a conserved MT-A70 enzyme domain of 177 aa,it has no signal peptide and transmembrane structural domain.METTL14is localized in the nucleus(69.60%)and has 40potential phosphorylation sites.The secondary structure of METTL14protein consists of irregularly coiled(51.10%),α-helix(33.11%),extended chain(11.84%)andβ-turn(3.95%)interconnections.METTL14interacts with methylation modification-related proteins and is mainly involved in the process of RNA methylation modification.The results of qRT-PCR showed that METTL14mRNA was expressed in all seven tissues of yak,with the highest expression in the longest dorsal muscle compared to adipose tissue(P<0.001).In temporal dimension,the expression of METTL14at 30months of age was more than that at 18 months of age(P<0.05).Further study revealed that METTL14expression showed an increasing trend at 0,4,8,12d during the differentiation of yak preadipocytes;the expression at 4,8d was significantly higher than that at 0d(P<0.05);the expression at 12dwas highly significantly higher than that at 0d(P<0.01).The results of this study indicate that METTL14,an acidic and water-soluble protein localized in the nucleus with a conserved structural domain of MT-A70,may play an important role in yak fat deposition in yaks.The present study
作者
马兰花
顾亚荣
张永峰
潘和平
阎萍
MA Lanhua;GU Yarong;ZHANG Yongfeng;PAN Heping;YAN Ping(College of Life Science and Engineering,North-west University for Nationalities,Lanzhou 730030,China;Key Laboratory of Yak Breeding Engineering of Gansu Province,Lanzhou Institute of Animal Husbandry and Veterinary Drug,Chinese Academy of Agricultural Sciences,Lanzhou 730030,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2022年第5期962-972,980,共12页
Chinese Journal of Veterinary Science
基金
甘肃省重点研发计划资助项目(17YF1NA067)
国家自然科学基金资助项目(31972561)
中国农业科学院牦牛资源与育种创新团队资助项目(CAASASTIP-2014-LIHPS-01)。
关键词
大通牦牛
甲基转移酶样蛋白14
生信分析
时空表达规律
Datong yak
methyltransferase-like protein 14
boinformatics analysis
spatiotemporal expression regularity
