JIA-2021-1319 ZNF2基因的启动子区一段314bp的SINE插入可能是与猪脂肪沉积相关的抑制因子

doi:10.1016/j.jia.2022.08.128

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (2): 526-536.DOI: 10.1016/j.jia.2022.08.128

JIA-2021-1319 ZNF2基因的启动子区一段314bp的SINE插入可能是与猪脂肪沉积相关的抑制因子

收稿日期:2021-08-16 接受日期:2022-01-22 出版日期:2023-02-20 发布日期:2022-01-22

A 314-bp SINE Insertion in the ZNF2 promoter region may act as a repressor related to regulation of fat deposition in pigs

GU Hao^1*, DU Zhan-yu^1*, Eduard MURANI², Enrico D’ALESSANDRO³, CHEN Cai¹, WANG Xiao-yan¹, MAO Jiu-de⁴, Klaus WIMMERS², SONG Cheng-yi¹

¹College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, P.R.China
²Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf 18196, Germany
³Department of Veterinary Science, Unit of Animal Production, University of Messina, Messina 98168, Italy
⁴Bond Life Science Center, University of Missouri, Columbia, MO, 65211, USA

Received:2021-08-16 Accepted:2022-01-22 Online:2023-02-20 Published:2022-01-22
About author:Received 16 August, 2021 Accepted 22 January, 2022 GU Hao, E-mail: guhao1213@outlook.com; DU Zhan-yu, E-mail: yuzhan.du@outlook.com; Correspondence SONG Cheng-yi, Tel: +86-514-87979034, E-mail: cysong@yzu.edu.cn * These authors contributed equally to this study.
Supported by:
This research was supported by the National Natural Science Foundation of China (32002146 and 31872977), the China Postdoctoral Science Foundation (2020M671630), the Jiangsu Postdoctoral Science Foundation of China (2021K221B) to Chen Cai, the Jiangsu Agriculture Science and Technology Innovation Fund, China [CX (19) 2016], and the Priority Academic Program Development of Jiangsu Higher Education Institutions and the High-end Talent Support Program of Yangzhou University, China to Song Chengyi.

摘要/Abstract

摘要：

反转录转座子是一种可以在基因组上移动的DNA片段，是引起遗传变异的重要来源，而基于其插入多态性可以用于开发分子标记。锌指蛋白(ZNFs)是真核生物中含量最丰富的蛋白质之一，其功能极其多样。本研究通过生物信息学方法，在6个锌指蛋白基因(ZNF2、ZNF3、ZNF7、ZNF8、ZNF10和ZNF12)中进行了反转录转座子插入多态性标记的预测，进而通过PCR实验进行验证，然后进一步研究ZNF2基因第1个内含子区的SINE转座子插入对基因活性的影响，借助双荧光素酶系统在3种细胞系（HeLa、PEF和PK15）中进行了检测分析，同时在大白猪群体中对这一位点开展了基因分型并分析了其对猪相关生产性能的影响。实验结果表明，上述ZNFs基因中存在6个反转录转座子多态性位点，它们分别是：ZNF2基因第1内含子中的SINE介导的1个多态位点和第3内含子中L1介导的1个多态位点;ZNF3基因5’侧翼区中SINE介导的1个多态位点和第2内含子中SINE介导的1个多态位点;ZNF7基因3’非翻译区中SINE介导的1个多态位点以及ZNF12基因第2内含子中L1介导的1个多态位点。在HeLa和PEF细胞中，ZNF2基因第1内含子区中的SINE插入极显著(P<0.01)的抑制了其核心启动子的活性，进而推测该位点的SINE可能是ZNF2基因的一种抑制因子，该SINE插入多态标记也显著(P<0.05)影响了大白猪的校正背膘厚，表现为该位点存在SINE插入的个体的校正背膘厚显著(P<0.05)高于无SINE插入的个体。综上所述，我们的数据表明，反转录转座子插入多态在这些锌指蛋白基因的遗传变异中扮演着重要的角色，ZNF2基因第1内含子区中的SINE插入多态标记有望成为在猪育种中进行脂肪沉积筛选的新型分子标记。

Abstract:

Retrotransposons, a type of DNA fragment that can mobilize itself on genome, can generate genetic variations and develop for molecular markers based on the insertion polymorphism. Zinc finger proteins (ZNFs) are among the most abundant proteins in eukaryotic animals, and their functions are extraordinarily diverse and particularly important in gene regulation. In the current study, bioinformatic prediction was performed to screen for retrotransposon insertion polymorphisms (RIPs) in six ZNF genes (ZNF2, ZNF3, ZNF7, ZNF8, ZNF10 and ZNF12). Six RIPs in these ZNFs, including one short interspersed nuclear element (SINE) RIP in intron 1 and one long interspersed nuclear element 1 (L1) RIP in intron 3 of ZNF2, one SINE RIP in 5´ flanking region and one SINE RIP in intron 2 of ZNF3, one SINE RIP in 3´ UTR of ZNF7 and one L1 RIP in intron 2 of ZNF12, were discovered and their presence was confirmed by PCR. The impact of the SINE RIP in the first intron of ZNF2, which is close to the core promoter of ZNF2, on the gene activity was investigated by dual-luciferase assay in three cell lines. Our results showed that the SINE insertion in the intron 1 of ZNF2 repressed the core promoter activity extremely significantly (P<0.01) in cervical cancer cells and porcine primary embryonic fibroblasts (HeLa and PEF), thus SINE may act as a repressor. This SINE RIP also significantly (P<0.05) affected the corrected back fat thickness in Yorkshire pigs. The corrected back fat thickness of individuals with SINE insertion in the first intron of ZNF2 was significantly (P<0.05) higher than that of individuals without SINE insertion. In summary, our data suggested that RIPs play important roles in the genetic variations of these ZNF genes and SINE RIP in the intron 1 of ZNF2 may provide a useful molecular marker for the screening of fat deposition in the pig breeding.

Key words: retrotransposon , ZNF2 , polymorphism , repressor , fat deposition

. JIA-2021-1319 ZNF2基因的启动子区一段314bp的SINE插入可能是与猪脂肪沉积相关的抑制因子[J]. Journal of Integrative Agriculture, 2023, 22(2): 526-536.

GU Hao, DU Zhan-yu, Eduard MURANI, Enrico D’ALESSANDRO, CHEN Cai, WANG Xiao-yan, MAO Jiu-de, Klaus WIMMERS, SONG Cheng-yi. A 314-bp SINE Insertion in the ZNF2 promoter region may act as a repressor related to regulation of fat deposition in pigs[J]. Journal of Integrative Agriculture, 2023, 22(2): 526-536.

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JIA-2021-1319 ZNF2基因的启动子区一段314bp的SINE插入可能是与猪脂肪沉积相关的抑制因子

A 314-bp SINE Insertion in the ZNF2 promoter region may act as a repressor related to regulation of fat deposition in pigs

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