大白猪ZnT基因家族生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2022.11.002

摘要/Abstract

摘要： 【目的】分析大白猪溶质载体30A （ZnT）基因家族成员的生物学特性，并检测其在猪不同组织中的表达水平，为研究ZnT基因家族调控锌的代谢提供科学依据。【方法】使用多种生物信息学软件对猪ZnT基因家族10个成员（ZnT1~ZnT10）进行序列分析，包括基序、保守结构域、跨膜结构域、进化树、蛋白相互作用分析；使用实时荧光定量PCR方法检测ZnT基因家族在6月龄大白猪心脏、肝脏、脾脏、肺脏、肾脏、卵巢、乳腺中的表达差异。【结果】 ZnT1~ZnT10基因定位到猪的8条染色体上，分子质量在41.61~85.30 ku之间；等电点介于6.27~9.30之间，其中ZnT6等电点最高为9.30，ZnT1等电点最低为6.27。ZnT基因家族中，除ZnT3外都属于稳定蛋白，除ZnT5、ZnT9外都含有6层跨膜结构域，除ZnT6、ZnT9外所含基序顺序均一致，Cation-efflux是ZnT基因家族共有的保守结构域。ZnT6与ZnT2、ZnT3、ZnT7、ZnT9蛋白关联程度最高，ZnT1与ZnT3蛋白关联程度最高，ZnT8与其他家族成员间关系较远。ZnT基因家族序列在哺乳动物进化过程中相对保守。ZnT基因家族的10个基因在大白猪肺脏中相对表达量较高，在心脏中相对表达量较低，除ZnT5、ZnT7、ZnT9外的ZnT基因家族的其他成员在肌肉中几乎不表达。【结论】 ZnT家族属于跨膜结构蛋白，所含基序顺序基本一致，Cation-efflux是ZnT基因家族共有的保守结构域。ZnT基因家族多数成员在大白猪肺脏中表达量最高，在心脏和肌肉中相对表达量较低。

关键词: 大白猪; ZnT基因家族; 生物信息学分析; 组织表达

Abstract: 【Objective】 The aim of this study was to analyze the biological characteristics of solute carrier 30A(ZnT) gene in Yorkshire pigs, and detect the expression of ZnT gene in different tissues of pigs, so as to provide a scientific basis for the study on the regulation of zinc metabolism by ZnT gene family.【Method】 Various bioinformatics software were used to analyze the sequence of ZnT gene family (ZnT1-ZnT10), including motif, conserved domain, transmembrane domain, evolutionary tree and protein interaction.Real-time quantitative PCR was used to detect the expression differences of ZnT gene family in heart, liver, spleen, lung, kidney, ovary and breast in Yorkshire pigs.【Result】 ZnT1-ZnT10 genes were located on 8 chromosomes of pigs, the molecular weight were 41.61-85.30 ku, and the isoelectric points were 6.27(ZnT1)-9.30(ZnT6).In the ZnT gene family, they all belonged to stable proteins except for ZnT3, contained 6-layer transmembrane domains except for ZnT5 and ZnT9, and the sequence of motifs were the same except for ZnT6 and ZnT9.Cation-efflux was a common conservative domain of ZnT gene family.ZnT6 and ZnT2, ZnT3, ZnT7 and ZnT9 proteins had the highest interrelationship, ZnT1 and ZnT3 proteins had the highest interrelationship, and ZnT8 had a distant relationship with other family members.The sequences of ZnT gene family were relatively conserved during mammalian evolution.The 10 genes of ZnT gene family had higher expression in lung and lower expression in heart of Yorkshire pigs, except for ZnT5, ZnT7 and ZnT9 genes, the other members of ZnT gene family were almost not expressed in muscle.【Conclusion】 ZnT family belonged to transmembrane structural proteins, and the motifs were basically in the same order.Cation-efflux was a common conserved domain of ZnT gene family.Most members of ZnT gene family had the highest expression in lung, and the low expression in heart and muscle of Yorkshire pigs.

Key words: Yorkshire pigs; ZnT gene family; bioinformatics analysis; tissue expression

中图分类号:

李月月, 吕冬玲, 刘笑笑, 田威龙, 梁靓, 陈奎蓉, 程锋, 兰干球, 梁晶. 大白猪ZnT基因家族生物信息学分析及组织表达研究[J]. 中国畜牧兽医, 2022, 49(11): 4119-4128.

LI Yueyue, LYU Dongling, LIU Xiaoxiao, TIAN Weilong, LIANG Liang, CHEN Kuirong, CHENG Feng, LAN Ganqiu, LIANG Jing. Bioinformatics Analysis and Tissue Expression of ZnT Gene Family in Yorkshire Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4119-4128.

参考文献

[1] HARA T, TAKEDA T, TAKAGISHI T, et al.Physiological roles of zinc transporters:Molecular and genetic importance in zinc homeostasis[J].The Journal of Physiological Sciences, 2017, 67(2):283-301.
[2] MARET W, LI Y.Coordination dynamics of zinc in proteins[J].Chemical Reviews, 2009, 109(10):4682-4707.
[3] SKALNY A, ASCHNER M, TINKOV A.Zinc[J].Advances in Food and Nutrition Research, 2021, 96:251-310.
[4] ANDREWS G K, WANG H, DEY S K, et al.Mouse zinc transporter 1 gene provides an essential function during early embryonic development[J].Genesis, 2004, 40(2):74-81.
[5] GOLAN Y, KAMBE T, ASSARAF Y G.The role of the zinc transporter SLC30A2/ZnT2 in transient neonatal zinc deficiency[J].Metallomics, 2017, 9:1352-1366.
[6] MCCORMICK N H, LEE S, HENNIGAR S R, et al.ZnT4(SLC30A4)-null ("lethal milk") mice have defects in mammary gland secretion and hallmarks of precocious involution during lactation[J].American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 2016, 310(1):R33-R40.
[7] NISHIDA K, HASEGAWA A, NAKAE S, et al.Zinc transporter ZnT5/SLC30A5 is required for the mast cell-mediated delayed-type allergic reaction but not the immediate-type reaction[J].Journal of Experimental Medicine, 2009, 206(6):1351-1364.
[8] KABLE M E, RIAZATI N, KIRSCHKE C P, et al.The ZnT7-null mutation has sex dependent effects on the gut microbiota and goblet cell population in the mouse colon[J].PLoS One, 2020, 15(9):e0239681.
[9] HARDY A B, WIJESEKARA N, GENKIN I, et al.Effects of high-fat diet feeding on ZnT8-null mice:Differences between β-cell and global knockout of ZnT8[J].American Journal of Physiology-Endocrinology and Metabolism, 2012, 302(9):E1084-E1096.
[10] HOJYO S, FUKADA T.Zinc transporters and signaling in physiology and pathogenesis[J].Archires of Biochemistry and Biophysics, 2016, 611:43-50.
[11] HUANG L P, TEPAAMORNDECH S.The SLC30 family of zinc transporters-A review of current understanding of their biological and pathophysiological roles[J].Molecular Aspects of Medicine, 2013, 34(2-3):548-560.
[12] JAGADISH S, HOWARD L, THATI G.SLC30A10:A typical presentation of gene mutation with hypermanganesemia[J].Seizures and Polycythemia, 2021, 16:100505.
[13] CONKLIN D S, MCMASTER J A, CULBERTSON M R, et al.A gene involved in cobalt accumulation in Saccharomyces cerevisiae[J].Molecular and Cellular Biology, 1992, 12(9):3678-3688.
[14] KAMBE T, NARITA H, YAMAGUCHI-IWAI Y, et al.Cloning and characterization of a novel mammalian zinc transporter, zinc transporter 5, abundantly expressed in pancreatic beta cells[J].Journal of Biological Chemistry, 2002, 277(21):19049-19055.
[15] AMANDA K, OMOTOLA G, KATHRYN K, et al.Evolutionary rate covariation identifies SLC30A9(ZnT9) as a mitochondrial zinc transporter[J].The Biochemical Journal, 2021, 478(17):3205-3220.
[16] 孟婕, 张贝, 杨晴, 等.鸡NLRs基因家族鉴定、系统进化及表达分析[J].中国家禽, 2022, 44(3):1-8. MENG J, ZHANG B, YANG Q, et al.Identification, systematic evolution and expression analysis of NLRs gene family in chicken[J].Chinese Poultry, 2022, 44(3):1-8.(in Chinese)
[17] PALMITER R D, COLE T B, FINDLEY S D.ZnT-2, a mammalian protein that confers resistance to zinc by facilitating vesicular sequestration[J].European Molecular Biology Organization, 1996, 15(8):1784-1791.
[18] 陈嫒, 谢良地.锌稳态、金属反应转录因子与肺高血压[J].中华高血压杂志, 2021, 29(8):728-732. CHEN A, XIE L G.Zinc steady-state, metal-responsive transcription factors and pulmonary hypertension[J].Chinese Journal of Hypertension, 2021, 29(8):728-732.(in Chinese)
[19] GOMEZ N, DAVICINO R, BIAGGIO V, et al.Overexpression of inducible nitric oxide synthase and cyclooxygenase-2 in rat zinc-deficient lung:Involvement of a NF-kappaB dependent pathway[J]. Nitric Oxide, 2006, 14(1):30-38.
[20] KAMBE T.Molecular architecture and function of ZnT transporters[J]. Current Topics in Membranes, 2012, 69:199-220.
[21] DEMPSKI R E.The cation selectivity of the ZIP transporters[J].Current Topics in Membranes, 2012, 69:221-245.
[22] KAMBE T, HASHIMOTO A, FUJIMOTO S.Current understanding of ZIP and ZnT zinc transporters in human health and diseases[J].Cellular and Molecular Life Sciences, 2014, 71(17):3281-3295.
[23] CHOWANADISAI W, LONNERDAL B, KELLEHER S L.Identification of a mutation in SLC30A2(ZnT-2) in women with low milk zinc concentration that results in transient neonatal zinc deficiency[J].Journal of Biological Chemistry, 2006, 281(51):39699-39707.
[24] LOPEZ V, FOOLAD F, KELLEHER S L.ZnT2-overexpression represses the cytotoxic effects of zinc hyper-accumulation in malignant metallothionein-null T47D breast tumor cells[J]. Cancer Letters, 2011, 304(1):41-51.
[25] HILDEBRAND M S, PHILLIPS A M, MULLEN S A, et al.Loss of synaptic Zn²⁺ transporter function increases risk of febrile seizures[J]. Scientific Reports, 2015, 5(1):178169.
[26] BRUGGER D, HANAUER M, ORTNER J, et al.The response of zinc transporter gene expression of selected tissues in a pig model of subclinical zinc deficiency[J]. The Journal of Nutritional Biochemistry, 2021, 90:108576.
[27] WILLEKENS J, RUNNELS L W.Impact of zinc transport mechanisms on embryonic and brain development[J].Nutrients, 2022, 14(12):2526.
[28] 臧猛.猪锌转运蛋白基因的克隆与组织分布[D].郑州:河南农业大学, 2009. ZANG M.Molecular cloning and distribution of of pig zinc transporter[D].Zhengzhou:Henan Agricultural University, 2009.(in Chinese)