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FBXL3 Gene Expression and Correlation Analysis Between Its Polymorphism and Seasonal Estrus in Sheep (Ovis aries) |
YANG Yang1, ZHONG Ying-Jie1, JIANG Yu3, CHU Ming-Xing1,*, LIU Qiu-Yue2,* |
1 Key Laboratory of Animal Genetics, Breeding and Reproduction of Agriculture and Rural Affairs/Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2 Institute of Genetics and Developmental Biology/Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China; 3 Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province/College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China |
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Abstract Seasonal estrus limits reproduction efficiency of sheep (Ovis aries), therefore, it is of great significance to explore key genes that affect the seasonal estrus of sheep. In order to explore the expression level and polymorphism of the F-box and leucine-rich repeat protein 3 (FBXL3) gene which was found by whole genome resequencing (WGRS) and its association with seasonal estrus in sheep (Ovis aries), the present study selected Sunite sheep as research object, qPCR was used to investigate the expression of FBXL3 gene in different tissues and the influence of SNP on its expression. SequenomMassARRAY® SNP was applied to detect polymorphism of g.52551820G>A locus of FBXL3 gene in 2 different estrus patterns (year-round estrus group and seasonal estrus group). Then correlation analysis was carried out with seasonal estrus traits of sheep. Meanwhile, bioinformatics methods were used to analyze the effects of mutation on mRNA secondary structure and protein properties. The results showed that FBXL3 gene was widely expressed in the tissues of Sunite sheep, and 3 genetypes of GG, GA, and AA existed in the locus of FBXL3 g.52551820G>A. The expression of GG genotype was significantly higher than AA genotype in liver (P<0.01), kidney (P<0.01), and adrenal gland (P<0.05). The G was the dominant allele in seasonal estrus breed, while A was the dominant allele in year-round estrus breed. The genotype frequency and allele frequency of this locus were extremely significant different between seasonal estrous and year-round estrous breeds (P<0.01). Population genetic analysis showed that the g.52551820G>A locus of FBXL3 gene was at low polymorphism (PIC<0.25) in Prairie Tibetan sheep and Dorper sheep, and was at moderate polymorphism (0.25<PIC<0.50) in Tan sheep, Sunite sheep, Small Tail Han sheep and Suffolk sheep. Moreover, bioinformatics analysis showed that the hydrophilicity of FBXL3 amino acid residues near the mutation site decreased slightly, and the phosphorylation of the protein also changed. Altogether, the mutation at g.52551820G>A locus of FBXL3 gene might be a potential molecular marker of seasonal estrus in sheep, which provides a basic reference for studying the molecular mechanism of seasonal estrus.
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Received: 27 April 2021
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Corresponding Authors:
*mxchu@263.net;qyliu@genetics.ac.cn
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[1] 李倩. 2013. mRNA二级结构与基因功能和基因必要性的关系[D]. 硕士学位论文, 西北农林科技大学, 导师: 陶士珩, pp. 2-5. (Li Q.2013. Correlations among mRNA secondary structure, genefunctions and essentiality[D]. Thesis for M.S., Northwest A&F University, Supervisor: Tao S H, pp. 2-5.) [2] 石文昊, 童梦莎, 李恺, 等. 2018. 基于质谱的磷酸化蛋白质组学: 富集、检测、鉴定和定量[J]. 生物化学与生物物理进展, 45(12): 1250-1258. (Shi W H, Tong M S, Li K, et al.2018. Phosphoproteomics based on mass spectrometry (MS): Enrichment, detection, assignment and quantification[J]. Progress in Biochemistry and Biophysics, 45(12): 1250-1258.) [3] 时广森. 2014. FBXL3对近日节律作用机制的研究[D]. 博士学位论文, 南京大学, 导师: 徐璎, pp. 36-46. (Shi G S.2014. The roles of FBXL3 in regulating circadian clock[D]. Thesis for Ph.D., Nanjing University, Supervisor: Xu Y, pp. 36-46.) [4] 文禹粱, 王翔宇, 郭晓飞, 等. 2019. 不同产羔数小尾寒羊BMP4基因表达及其错义突变与产羔数关联分析[J]. 农业生物技术学报, 27(01): 80-88. (Wen Y L, Wang X Y, Guo X F, et al.2019. BMP4 gene expression in small tail han sheep (Ovis aries) with different litter size and association analysis between its missense mutation and litter size[J]. Chinese Journal of Agricultural Biotechnology, 27(01): 80-88.) [5] 钟英杰, 向光明, 狄冉, 等. 2020a. FBXL家族在哺乳动物生物节律中调控作用的研究进展[J]. 畜牧兽医学报, 51(02): 217-226. (Zhong Y J, Xiang G M, Di R, et al.2020a. Advances in the regulation of FBXL family in mammalian biorhythms[J]. Chinese Journal of Animal and Veterinary Sciences, 51(02): 217-226.) [6] 钟英杰, 向光明, 贺小云, 等. 2020b. FBXL3和FBXL21基因表达与苏尼特羊季节性发情关系初探[J]. 中国畜牧杂志, 56(12): 40-44. (Zhong Y J, Xiang G M, He X Y, et al.2020b. Preliminary study on the relationship between FBXL3 and FBXL21 gene expression and seasonal estrus in Sunite sheep[J]. Chinese Journal of Animal Science, 56(12): 40-44.) [7] 周梅, 曹晓涵, 贺小云, 等. 2018. 绵羊FSTL1基因组织表达及其多态性与产羔数之间的关系[J]. 农业生物技术学报, 26(05): 801-810. (Zhou M, Cao X H, He X Y, et al.2018. Tissue expression and polymorphism of sheep (Ovis aries) FSTL1 gene and their association with litter size[J]. Chinese Journal of Agricultural Biotechnology, 26(05): 801-810.) [8] Anand S N, Maywood E S, Chesham J E, et al.2013. Distinct and separable roles for endogenous CRY1 and CRY2 within the circadian molecular clockwork of the suprachiasmatic nucleus, as revealed by the Fbxl3Afh mutation[J]. The Journal of Neuroscience, 33(17): 7145-7153. [9] Ansar M, Paracha S A, Serretti A, et al.2019. Biallelic variants in FBXL3 cause intellectual disability, delayed motor development and short stature[J]. Human Molecular Genetics, 28(06): 972-979. [10] Busino L, Bassermann F, Maiolica A, et al.2007. SCFFbxl3 controls the oscillation of the circadian clock by directing the degradation of cryptochrome proteins[J]. Science, 316(5826): 900-904. [11] Chemineau P, Guillaume D, Migaud M, et al.2008. Seasonality of reproduction in mammals: Intimate regulatory mechanisms and practical implications[J]. Reproduction in Domestic Animals, 43(Suppl 2): 40-47. [12] Correia S P, Chan A B, Vaughan M, et al.2019. The circadian E3 ligase complex SCFFBXL3+CRY targets TLK2[J]. Science Report, 9(01): 198. [13] Dai Y X, Qiu M K, Wang S Q, et al.2020. lncRNA CASC2 suppresses the growth of hemangioma cells by regulating miR-18a-5p/FBXL3 axis[J]. Journal of Biological Regulators and Homeostatic Agents, 34(01): 49-56. [14] Dardente H, Mendoza J, Fustin J M, et al.2008. Implication of the F-box protein FBXL21 in circadian pacemaker function in mammals[J]. PLOS ONE, 3(10): e3530. [15] Di R, He J N, Song S H, et al.2014. Characterization and comparative profiling of ovarian microRNAs during ovine anestrus and the breeding season[J]. BMC Genomics, 15(01): 899. [16] Godinho S I, Maywood E S, Shaw L, et al.2007. The after-hours mutant reveals a role for Fbxl3 in determining mammalian circadian period[J]. Science, 316(5826): 897-900. [17] Goldman B D.1999. The circadian timing system and reproduction in mammals[J]. Steroids, 64(09): 679-685. [18] Guh Y J, Tamai T K, Yoshimura T.2019. The underlying mechanisms of vertebrate seasonal reproduction[J]. Proceedings of the Japan Academy, Series B, Physical and Biological Sciences, 95(07): 343-357. [19] Helfer G, Barrett P, Morgan P J.2019. A unifying hypothesis for control of body weight and reproduction in seasonally breeding mammals[J]. Journal of Neuroendocrinology, 31(03): e12680. [20] Huber A L, Papp S J, Chan A B, et al.2016. CRY2 and FBXL3 cooperatively degrade c-MYC[J]. Molecullar Cell, 64(04): 774-789. [21] Ikegami K, Yoshimura T.2012. Circadian clocks and the measurement of daylength in seasonal reproduction[J]. Molecullar and Cellular Endocrinology, 349(01): 76-81. [22] Julius A A, Yin J, Wen J T.2019. Time optimal entrainment control for circadian rhythm[J]. PLOS ONE, 14(12): e0225988. [23] La Y F, He X Y, Zhang L P.2020. Comprehensive analysis of differentially expressed profiles of mRNA, lncRNA, and circRNA in the uterus of seasonal reproduction sheep[J]. Genes (Basel), 11(03): 301. [24] Lincoln G, Messager S, Andersson H, et al.2002. Temporal expression of seven clock genes in the suprachiasmatic nucleus and the pars tuberalis of the sheep: Evidence for an internal coincidence timer[J]. Proceedings of the National Academy of Sciences of the USA, 99(21): 13890-13895. [25] Liu Z J, Qian M X, Tang X L, et al.2019. SIRT7 couples light-driven body temperature cues to hepatic circadian phase coherence and gluconeogenesis[J]. Nature Metabolism, 01(11): 1141-1156. [26] Mohawk J A, Green C B, Takahashi J S.2012. Central and peripheral circadian clocks in mammals[J]. Annual Review of Neuroscience, 35: 445-462. [27] Nakayama T, Yoshimura T.2018. Seasonal rhythms: The role of thyrotropin and thyroid hormones[J]. Thyroid, 28(01): 4-10. [28] Nishiwaki-Ohkawa T, Yoshimura T.2016. Molecular basis for regulating seasonal reproduction in vertebrates[J]. Journal of Endocrinology, 229(03): R117-R127. [29] Pan Z Y, Li S D, Liu Q Y, et al.2018. Whole-genome sequences of 89 Chinese sheep suggest role of RXFP2 in the development of unique horn phenotype as response to semi-feralization[J]. GigaScience, 7: 1-15. [30] Parico G C G, Partch C L.2020. The tail of cryptochromes: An intrinsically disordered cog within the mammalian circadian clock[J]. Cell Communication and Signaling, 18(01): 182. [31] Pittendrigh C S.1972. Circadian surfaces and the diversity of possible roles of circadian organization in photoperiodic induction[J]. Proceedings of the National Academy of Sciences of the USA, 69(09): 2734-2737. [32] Rose J, Kraft T, Brenner B, et al.2020. Hypertrophic cardiomyopathy MYH7 mutation R723G alters mRNA secondary structure[J]. Physiological Genomics, 52(01): 15-19. [33] Saran A R, Kalinowska D, Oh S, et al.2018. JMJD5 links CRY1 function and proteasomal degradation[J]. PLOS Biology, 16(11): e2006145. [34] Sato T K, Yamada R G, Ukai H, et al.2006. Feedback repression is required for mammalian circadian clock function[J]. Nature Genetics, 38(03): 312-319. [35] Shi G S, Xing L J, Liu Z W, et al.2013. Dual roles of FBXL3 in the mammalian circadian feedback loops are important for period determination and robustness of the clock[J]. Proceedings of the National Academy of Sciences of the USA, 110(12): 4750-4755. [36] Siepka S M, Yoo S H, Park J, et al.2007. Circadian mutant overtime reveals F-box protein FBXL3 regulation of cryptochrome and period gene expression[J]. Cell, 129(05): 1011-1023. [37] Wang D Z, Han X, Li C, et al.2019. FBXL3 is regulated by miRNA-4735-3p and suppresses cell proliferation and migration in non-small cell lung cancer[J]. Pathology, Research and Practice, 215(02): 358-365. [38] Yasuo S, Watanabe M, Tsukada A, et al.2004. Photoinducible phase-specific light induction of Cry1 gene in the pars tuberalis of Japanese quail[J]. Endocrinology, 145(04): 1612-1616. [39] Yumimoto K, Muneoka T, Tsuboi T, et al.2013. Substrate binding promotes formation of the Skp1-Cul1-Fbxl3 (SCFFbxl3) protein complex[J]. Journal of Biological Chemistry, 288(45): 32766-32776. [40] Zhai M J, Xie Y F, Liang H H, et al.2018. Comparative profiling of differentially expressed microRNAs in estrous ovaries of Kazakh sheep in different seasons[J]. Gene, 664: 181-191. |
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