b-Boule基因5′调控序列的克隆与睾丸组织DMR甲基化分析

doi:10.3864/j.issn.0578-1752.2011.18.017

Abstract

Abstract: 【Objective】 In order to reveal b-Boule gene expression and regulation, and to supply evidence for apparent genetic mechanism of cattle-yak male sterile, the 5′-flanking regulatory sequence of b-Boule, and the difference of b-Boule gene DMR methylation status in testes among cattle, yaks and cattle-yaks were studied. 【Method】 The 5′-flanking regulatory sequence of b-Boule in yak was cloned and sequenced by the PCR amplification, the 5´-flanking regulatory sequence of b-Boule was analyzed using bioinformatics, and the DMR methylation status of b-Boule gene in testes among cattle, yaks and cattle-yaks were detected using sodium bisulfite sequencing. 【Result】 The 5′-flanking regulatory sequence length of b-Boule is 1 352 bp, and there is a CpG island. The core promoter region contains SP1 methylation-sensitive site. The methylation level of b-Boule gene DMR in cattle-yaks (17.78%) was significantly greater than that in cattle (6.94%) and yaks (7.50%) (P＜0.01). The difference of the methylation level is obviously, especially the CpG site’s 33-35. 【Conclusion】 The methylation level of yak b-Boule gene DMR is significantly greater than that of cattle-yak. Combined with the previous study results about mRNA expression and histological observation, it was speculated that the DMR methylation played a key role in the expression and regulation of b-Boule, and it might arrest meiotic process of spermatogenesis by its mRNA expression inhibition through the high methylation level of b-Boule gene DMR in cattle-yak.

Key words: yak, cattle-yak, b-Boule gene, 5′regulation sequence, methylation

LI Ming-Gui, XU Hong-Tao, LI Yin-Xia, YU Sha-Li, ZHAO Xing-Bo, PAN Zeng-Xiang, ZHU Xiang, XIE Zhuang, LI Qi-Fa. Cloning and Testicular DMR Methylation Analysis of b-Boule Gene 5′-Flanking Regulation Regions[J].Scientia Agricultura Sinica, 2011, 44(18): 3859-3867.

References

[1]Shah C, Vangompel M J, Naeem V, Chen Y, Lee T, Angeloni N, Wang Y, Xu E Y. Widespread presence of human BOULE homologs among animals and conservation of their ancient reproductive function. PLoS Genetics, 2010, 6(7): e1001022.

[2]Reynolds N, Cooke H J. Role of the DAZ genes in male fertility. Reproductive Biomedicine Online, 2005, 10: 72-80.

[3]Collier B, Gorgoni B, Loveridge C, Cooke H J, Gray N K. The DAZL family proteins are PABP-binding proteins that regulate translation in germ cells. The EMBO Journal, 2005, 24: 2656-2666.

[4]Kee K, Angeles V T, Flores M, Nguyen H N, Reijo Pera R A. Human DAZL, DAZ and BOULE genes modulate primordial germ-cell and haploid gamete formation. Nature, 2009, 462: 222-225.

[5]Xu E Y, Moore F L, Reijo Pera R A. A gene family required for human germ cell development evolved from an ancient meiotic gene conserved in metazoan. Proceeding of the National Academy of Sciences of the United States of America, 2001, 98(13): 7414-7419.

[6]Luetjens C M, Xu E Y, Rejo Pera R A, Kamischke A, Nieschlag E, Gromoll J. Association of meiotic arrest with lack of BOULE protein expression in infertile men. Journal of Clinical Endocrinology Metabolism, 2004, 89: 1926-1933.

[7]屈旭光, 李齐发, 刘振山, 董丽艳, 李新福, 郝称莉, 谢庄. 牦牛、犏牛睾丸组织中SYCP3基因mRNA表达水平研究. 畜牧兽医学报, 2008, 39: 1132-1136.

Qu X G, Li Q F, Liu Z S, Dong L Y, Li X F, Hao C L, Xie Z. The study on the expression level of SYCP3 mRNA in yak an cattle-yak tesits. Acta Veterinaria et Zootechnica Sinica, 2008, 39: 1132-1136. (in Chinese)

[8]Xu E Y, Lee D F, Klebes A, Turek P J, Kornberg T B, Reijo Pera R A. Human BOULE gene rescues meiotic defects in infertile flies. Human Molecular Genetics, 2003, 12: 169-175.

[9]Tung J Y, Luetjens C M, Wistuba J, Xu E Y, Reijo Pera R A, Gromoll J. Evolutionary comparison of the reproductive genes, DAZL and BOULE, in primates with and without DAZ. Development Genes and Evolution, 2006, 216: 158-168.

[10]Xu H Y, Li Z D, Li M Y, Wang L, Hong Y H. Boule is present in fish and bisexually expressed in adult and embryonic germ cells of medaka. PLoS One, 2009, 4(6): e6097.

[11]Kuo P L, Wang S T, Lin Y M, Lin Y H, Teng Y N, Hsu C C. Expression profiles of the DAZ gene family in human testis with and without spermatogenic failure. Fertility and Sterility, 2004, 81: 1034-1040.

[12]Eberhart C G, Maines J Z, Wasserman S A. Meiotic cell cycle requirement for a fly homologue of human deleted in Azoospermia. Nature, 1996, 381: 783-785.

[13]VanGompel M J W, Xu E Y. A novel requirement in mammalian spermatid differentiation for the DAZ-family protein Boule. Human Molecular Genetics, 2010, 19: 2360-2369.

[14]Truong B N, Moses E K, Armes J E, Venter D J, Baker H W G. Searching for candidate genes for male infertility. Asian Journal of Andrology, 2003, 5: 137-147.

[15]Zhang Q B, Li Q F, Li J H, Li X F, Liu Z S, Song D W, Xie Z. b-DAZL: A novel gene in bovine spermatogenesis. Progress in Natural Science, 2008, 18: 1209-1218.

[16]Zhang Q B, Li J H, Li Q F, Li X F, Liu Z S, Song D W, Xie Z. Cloning and characterization of the gene encoding the bovine BOULE protein. Molecular Genetics and Genomics, 2009, 281: 67-75

[17]李新福, 李齐发, 李隐侠, 张庆波, 徐洪涛, 马志杰, 钟金城, 谢庄. 黄犏牛与黄牛b-Boule基因编码区序列、结构、表达模式和睾丸组织表达水平的差异分析. 自然科学进展, 2009, 19(10): 1042-1048.

Li X F, Li Q F, Li Y X, Zhang Q B, Xu H T, Ma Z J, Zhong J C, Xie Z. Progress in Nature Science, 2009, 19(10): 1042-1048. (in Chinese)

[18]Maatouk D M, Kellam L D, Mann M R W, Lei H, Li E, Bartolomei M S, Resnick J L. DNA methylation is a primary mechanism for silencing postmigratory primordial germ cell genes in both germ cell and somatic cell lineages. Development, 2006, 133: 3411-3418.

[19]Haston K M, Tung J Y, Reijo Pera R A. Dazl functions in maintenance of pluripotency and genetic and epigenetic programs of differentiation in mouse primordial germ cells in vivo and in vitro. PLoS One, 2009, 4(5): e5654.

[20]Linher K, Cheung Q, Baker P, Bedecarrats G, Shiota K, Li J. An epigenetic mechanism regulates germ cell-specific expression of the porcine deleted in azoospermia-like (DAZL) gene. Differentiation, 2009, 77: 335-349.

[21]Navarro-Costa P, Nogueira P, Carvalho M, Leal F, Cordeiro I, Calhaz-Jorge C, Gonçalves J, Plancha C E. Incorrect DNA methylation of the DAZL promoter CpG island associates with defective human sperm. Human Reproduction, 2010, 25(10): 2647-2654.

[22]李齐发, 李隐侠, 张庆波, 刘振山, 屈旭光, 谢庄. DAZ家族新成员BOULE蛋白的结构与功能. 中国生物化学与分子生物学报, 2007, 23(10): 823-828.

Li Q F, Li Y X, Zhang Q B, Liu Z S, Qu X G, Xie Z. Structure and function of boule protein, a new member of the human DAZ family. Chinese Journal of Biochemistry and Molecular Biology, 2007, 23(10): 823-828. (in Chinese)

[23]Mancini D N, Singh S M, Archer T K, Rodenhiser D I. Site-specific DNA methylation in the neurofibromatosis (NF1) promoter interferes with binding of CREB and SP1 transcription factors. Oncogene, 1999, 18: 4108-4119.

[24]Boumber Y A, Kondo Y, Chen X Q, Shen L L, Guo Y, Tellez C, Estécio M R H, Ahmed S, Issa J P J. An Sp1/Sp3 binding polymorphism confers methylation protection. PLoS Genetics, 2008, 4: e1000162.

[25]Walker W H, Habener J F. Role of transcription factors CREB and CREM in cAMP-regulated transcription during spermatogenesis. Trends in Endocrinology and Metabolism, 1996, 7: 133-138.

[26]Jones P A, Takai D. The role of DNA methylation in mammalian epigenetics. Science, 2001, 293: 1068-1070.

[27]Lorincz M C, Dickerson D R, Schmitt M, Groudine M. Intragenic DNA methylation alters chromatin structure and elongation efficiency in mammalian cells. Nature Structural and Molecular Biology, 2004, 11: 1068-1075.

[28]Bauer A P, Leikam D, Krinner S, Notka F, Ludwig C, Längst G, Wagner R. The impact of intragenic CpG content on gene expression. Nucleic Acids Research, 2010, 38: 3891-3908.

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Cloning and Testicular DMR Methylation Analysis of b-Boule Gene 5′-Flanking Regulation Regions

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