伊犁马ACTN3基因外显子多态性研究

doi:10.11843/j.issn.0366-6964.2018.06.023

摘要/Abstract

摘要：

旨在鉴定新疆伊犁马ACTN3基因外显子多态位点及其单倍型，分析不同单倍型对mRNA表达量和蛋白质结构的影响，为进一步探究其基因型与运动性能的关联奠定基础。以38匹伊犁马为研究对象，通过直接测序法筛选ACTN3基因多态性位点并进行基因型分析，并通过haploview 4.2软件进行单倍型分析；利用在线软件（RNAfold和sopm）进行生物信息学分析。结果表明，在伊犁马ACTN3基因的外显子中共发现了8个多态位点，其中T9059G为错义突变，其余均为同义突变；A6206G和A11517G之间存在强的连锁，其余各突变间也存在一定程度的连锁关系；构建出的14种单倍型中，单倍型H1（GGATCCCG）为优势单倍型，各单倍型使得ACTN3基因mRNA二级结构和最小自由能发生改变，且T9059G使得蛋白质二级结构改变。结果提示，T9059G突变位点影响ACTN3蛋白质的二级结构，可能是对伊犁马运动性能产生影响的重要功能位点。

Abstract:

The aims of this study were to identify genetic variants and construct haplotypes of polymorphic sites of ACTN3 gene exon in Yili horse, and determine different haplotypes effects on the mRNA expression level and protein secondary stucture of ACTN3, which would provide a theoretical basis for further researching the influence of the gene expression on athletic performance. Direct sequencing was taken to detect the polymorphic sites of of ACTN3 of 38 Yili horse and analyze its genotypes. The haplotypes and bioinformation were analyzed by haploview 4.2 and online software (RNAfold and sopm), respectively. A total of 8 SNPs were identified from the ACTN3 gene exon, only T9059G was the missense mutant, and the other 7 were synonymous mutants. Based on the 8 SNPs, linkage analysis showed that the strong linkage between A6206G and A11517G, and the different linkage relationship among the other SNPs. There was 14 haplotypes by the 8 SNPs, H1(GGATCCCG) was the dominant haplotype, and the different haplotypes could cause the change of mRNA secondary structure and minimum free energy of ACTN3 gene. The missense mutation of T9059G changed the secondary structure of ACTN3 protein. The result indicate that the T9059G is one of the improtant functional sites affecting athletic performance, which affect the secondary structure of ACTN3 protein.

中图分类号:

S821.2

王建文, 李林玲, 王镜力, 孟军, 曾亚琦, 姚新奎, 辛雅莉. 伊犁马ACTN3基因外显子多态性研究[J]. 畜牧兽医学报, 2018, 49(6): 1299-1306.

WANG Jian-wen, LI Lin-ling, WANG Jing-li, MENG Jun, ZENG Ya-qi, YAO Xin-kui, XIN Ya-li. Analysis on ACTN3 Gene Exon Polymorphism of Yili Horse[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1299-1306.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.06.023

https://www.xmsyxb.com/CN/Y2018/V49/I6/1299

参考文献

[1] BEGGS A H,BYERS T J,KNOLL J H,et al.Cloning and characterization of two human skeletal muscle alpha-actinin genes located on chromosomes 1 and 11[J].J Biol Chem,1992,267(13):9281-9288.
[2] DEL COSO J,VALERO M,SALINERO J J,et al.ACTN3 genotype influences exercise-induced muscle damage during a marathon competition[J].Eur J Appl Physiol,2017,117(3):409-416.
[3] MARCHLER-BAUER A,LU S,ANDERSON J B,et al.CDD:a Conserved Domain Database for the functional annotation of proteins[J].Nucleic Acids Res,2011,39(D1):D225-D229.
[4] GINEVICENE V,JAKAITIENE A,AKSENOV M O,et al.Association analysis of ACE,ACTN3 and PPARGC1A gene polymorphisms in two cohorts of European strength and power athletes[J].Biol Sport,2016,33(3):199-206.
[5] MIN S K,LIM S T,KIM C S.Association of ACTN3 polymorphisms with BMD,and physical fitness of elderly women[J].J Phys Ther Sci,2016,28(10):2731-2736.
[6] GARTON F C,NORTH K N.The Effect of heterozygosity for the ACTN3 null allele on human muscle performance[J].Med Sci Sport Exer,2016,48(3):509-520.
[7] The PLoS Genetics Staff.Correction:altered Ca²⁺ kinetics associated with α-actinin-3 deficiency may explain positive selection for ACTN3 null allele in human evolution[J].PLoS Genet,2015,11(6):e1005312.
[8] HEAD S I,CHAN S,HOUWELING P J,et al.Altered Ca²⁺ kinetics associated with α-actinin-3 deficiency may explain positive selection for ACTN3 null allele in human evolution[J].PLoS Genet,2015,11(2):e1004862.
[9] GU J,MACHUGH D E,MCGIVNEY B A,et al.Association of sequence variants in CKM (creatine kinase,muscle) and COX4I2(cytochrome c oxidase,subunit 4,isoform 2) genes with racing performance in thoroughbred horses[J].Equine Vet J,2010(38):569-575.
[10] MATA X,VAIMAN A,DUCASSE A,et al.Genomic structure,polymorphism and expression of the horse alpha-actinin-3 gene[J].Gene,2012,491(1):20-24.
[11] THOMAS K C,HAMILTON N A,NORTH K N,et al.Sequence analysis of the equine ACTN3 gene in Australian horse breeds[J].Gene,2014,538(1):88-93.
[12] 赵启南,芒来,白东义,等.蒙古马高负荷运动训练前后转录组差异表达分析[J].畜牧兽医学报,2017,48(6):1007-1016.
ZHAO Q N,MANG L,BAI D Y,et al.Mongolian horses transcriptome differential expression analysis before and after a high load exercise training[J].Acta Veterinaria et Zootechnica Sinica,2017,48(6):1007-1016.(in Chinese)
[13] 姚新奎,韩国才.马生产管理学[M].北京:中国农业大学出版社,2008.
YAO X K,HAN G C.Horse production and management[M].Beijing:China Agricultural University Press,2008.(in Chinese)
[14] 中华人民共和国农业部. NY/T 2831-2015伊犁马[S].北京:中国农业出版社,2015.
The Ministry of Agriculture of the People's Republic of China.NY/T 2831-2015 Yili Horse[S].Beijing:China Agriculture Press,2015.(in Chinese)
[15] DE MORAES V N,FERRARI G D,CHIARATTO T,et al.Association of genetic polymorphisms with physical capacities and body composition in older women[J].Rev Bras Cineantropom Desempenho Hum,2016,18(1):11-19.
[16] BAUMERT P,LAKE M J,STEWART C E,et al.Genetic variation and exercise-induced muscle damage:implications for athletic performance,injury and ageing[J].Eur J Appl Physiol,2016,116(9):1595-1625.
[17] PICKERING C,KIELY J.ACTN3:more than just a gene for speed[J].Front Physiol,2017,8:1080.
[18] DIONÍSIO T J,THIENGO C R,BROZOSKI D T,et al.The influence of genetic polymorphisms on performance and cardiac and hemodynamic parameters among Brazilian soccer players[J].Appl Physiol Nutri Metab,2017,42(6):596-604.
[19] KIKUCHI N,MIYAMOTO-MIKAMI E,MURAKAMI H,et al.ACTN3 R577X genotype and athletic performance in a large cohort of Japanese athletes[J].Eur J Sport Sci,2016,16(6):694-701.
[20] PAPADIMITRIOU I D,LUCIA A,PITSILADIS Y P,et al.ACTN3,R577X and ACE I/D gene variants influence performance in elite sprinters:a multi-cohort study[J].Bmc Genomics,2016,17:285.
[21] SETO J T,QUINLAN K G R,LEK M,et al.ACTN3 genotype influences muscle performance through the regulation of calcineurin signaling[J].J Clin Invest,2013,123(10):4255-4263.
[22] EBASHI S,EBASHI F.α-actinin,a new structural protein from striated muscle:I.Preparation and action on actomyosin-ATP interaction[J].J Biochem,1965,58(1):7-12.
[23] NORTH K.Why is α-actinin-3 deficiency so common in the general population? The evolution of athletic performance[J].Twin Res Hum Genet,2008,11(4):384-394.
[24] LIN Z,HIJIKATA T,ZHANG Z,et al.Dispensability of the actin-binding site and spectrin repeats for targeting sarcomeric α-actinin into maturing Z bands in vivo:implications for in vitro binding studies[J].Dev Biol,1998,199(2):291-308.
[25] YANG N,GARTON F,NORTH K.α-actinin-3 and performance[J].Med Sport Sci,2009,54:88-101.
[26] YIN H L,JANMEY P A.Phosphoinositide regulation of the actin cytoskeleton[J].Annu Rev Physiol,2003,65(1):761-789.
[27] BARON M D,DAVISON M D,JONES P,et al.The sequence of chick alpha-actinin reveals homologies to spectrin and calmodulin[J].J Biol Chem,1987,262(36):17623-17629.
[28] DJINOVIC-CARUGO K,GAUTEL M,YLÄNNE J,et al.The spectrin repeat:a structural platform for cytoskeletal protein assemblies[J].FEBS Lett,2002,513(1):119-123.
[29] HOGARTH M W,HOUWELING P J,THOMAS K C,et al.Evidence for ACTN3 as a genetic modifier of Duchenne muscular dystrophy[J].Nat Commun,2017,8:14143.
[30] TANG J H,TAYLOR D W,TAYLOR K A.The three-dimensional structure of α-actinin obtained by cryoelectron microscopy suggests a model for Ca²⁺-dependent actin binding 1[J].J Mol Biol,2001,310(4):845-858.
[31] GARTON F C,SETO J T,QUINLAN K G R,et al.α-Actinin-3 deficiency alters muscle adaptation in response to denervation and immobilization[J].Hum Mol Genet,2014,23(7):1879-1893.
[32] 轩俊丽,马晓萌,王慧华,等.绵羊季节性繁殖相关基因TSHR外显子多态性研究[J].畜牧兽医学报,2016,47(7):1342-1353.
XUAN J L,MA X M,WANG H H,et al.Study on exon polymorphism of seasonal breeding related gene TSHR in sheep[J].Acta Veterinaria et Zootechnica Sinica,2016,47(7):1342-1353.(in Chinese)