Association of TNP2 Gene Polymorphisms of the bta-miR-154 Target Site with the Semen Quality Traits of Chinese Holstein Bulls.docVIP

AssociationofTNP2GenePolymorphismsofthebta- miR-154TargetSitewiththeSemenQualityTraitsof ChineseHolsteinBulls QingGao1,2.,ZhihuaJu1.,YanZhang1.,JinmingHuang1,XiaojianZhang1,ChaoQi1,JianbinLi1, JifengZhong1,GuorongLi2*,ChangfaWang1* 1DairyCattleResearchCenter,ShandongAcademyofAgriculturalScience,Jinan,PRChina,2CollegeofLifeScience,ShandongNormalUniversity,Jinan,PRChina Abstract Transition protein 2 (TNP2) participates in removing nucleohistones and the initial condensation of spermatid nucleus during spermiogenesis. This study investigated the relationship between the variants of the bovine TNP2 gene and the semenqualitytraitsofChineseHolsteinbulls.Wedetectedthreesinglenucleotidepolymorphisms(SNPs)oftheTNP2gene in 392 Chinese Holstein bulls, namely, g.269 G.A (exon 1), g.480 C.T (intron 1), and g.1536 C.T (39-UTR). Association analysisshowedthatthesemenqualitytraitsoftheChineseHolsteinbullswassignificantlyaffectedbythethreeSNPs.The bullswiththehaplotypiccombinationsH6H4,H6H6,andH6H8hadhigherinitialsemenmotilitythanthosewiththeH7H8 andH8H4haplotypiccombinations(P,0.05).SNPsinthemicroRNA(miRNA)bindingregionoftheTNP2gene39-UTRmay havecontributed tothe phenotypic differences. The phenotypic differencesarecaused by thealtered expressionof the miRNAsandtheirtargets.Bioinformaticsanalysispredictedthattheg.1536C.TsiteintheTNP239-UTRislocatedinthe bta-miR-154 binding region. The quantitative real-time polymerase chain reaction results showed that the TNP2 mRNA relativeexpressioninbullswiththeCTandCCgenotypeswassignificantlyhigherthanthosewiththeTTgenotype(P,0.05) intheg.1536C.Tsite.Theluciferaseassayalsoindicatedthatbta-miR-154directlytargetsTNP2inamurineLeydigcell tumorcellline.TheSNPg.1536C.TintheTNP239-UTR,whichalteredthebindingofTNP2withbta-miR-154,wasfoundto beassociatedwiththesemenqualitytraitsofChineseHolsteinbulls. Citation:GaoQ,JuZ,ZhangY,HuangJ,ZhangX,etal.(2014)AssociationofTNP2GenePolymorphismsofthebta-miR-154TargetSitewiththeSemenQuality TraitsofChineseHolsteinBu