Splicing-related single nucleotide polymorphism of RAB, member of RAS oncogene family like 2B (RABL2B) jeopardises semen quality in Chinese Holstein bulls
Xiuge Wang A B , Xiaohui Cui B , Yan Zhang A B , Haisheng Hao A , Zhihua Ju B , Deyu Liu C , Qiang Jiang B , Chunhong Yang B , Yan Sun B , Changfa Wang B , Jinming Huang A B D and Huabin Zhu A D
+ Author Affiliations
- Author Affiliations
A Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan Western Road, Haidian District, Beijing 100193, PR China.
B Dairy Cattle Research Center, Shandong Academy of Agricultural Sciences, No. 159 North of Industry Road, Jinan, Shandong 250131, PR China.
C Station of Livestock and Poultry Improvement, Guangxi Zhuang Autonomous Region, No. 24 Yongwu Road, Nanning 530001, PR China.
D Corresponding authors. Emails: huangjinm@sina.com; zhuhuabin@caas.cn
Reproduction, Fertility and Development 29(12) 2411-2418 https://doi.org/10.1071/RD17111
Submitted: 5 December 2016 Accepted: 18 April 2017 Published: 30 May 2017
Abstract
RAB, member of RAS oncogene family like 2B (RABL2B) is a member of a poorly characterised clade of the RAS GTPase superfamily, which plays an essential role in male fertility, sperm intraflagellar transport and tail assembly. In the present study, we identified a novel RABL2B splice variant in bovine testis and spermatozoa. This splice variant, designated RABL2B-TV, is characterised by exon 2 skipping. Moreover, a single nucleotide polymorphism (SNP), namely c.125G>A, was found within the exonic splicing enhancer (ESE) motif, indicating that the SNP caused the production of the RABL2B-TV aberrant splice variant. This was demonstrated by constructing a pSPL3 exon capturing vector with different genotypes and transfecting these vectors into murine Leydig tumour cell line (MLTC-1) cells. Expression of the RABL2B-TV transcript was lower in semen from high- versus low-performance bulls. Association analysis showed that sperm deformity rate was significantly lower in Chinese Holstein bulls with the GG or GA genotype than in bulls with the AA genotype (P < 0.05). In addition, initial sperm motility was significantly higher in individuals with the GG or GA genotype than in individuals with the AA genotype (P < 0.05). The findings of the present study suggest that the difference in semen quality in bulls with different RABL2B genotypes is generated via an alternative splicing mechanism caused by a functional SNP within the ESE motif.
Additional keywords: functional single nucleotide polymorphism, splice variant.
References
Adhiambo, C., Blisnick, T., Toutirais, G., Delannoy, E., and Bastin, Pl. (2009). A novel function for the atypical small G protein Rab-like 5 in the assembly of the trypanosome flagellum. J. Cell Sci. 122, 834–841.| A novel function for the atypical small G protein Rab-like 5 in the assembly of the trypanosome flagellum.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXksFWqsrc%3D&md5=e43c49c8e2aec2cc4d963ea8f51c2769CAS |
Allan, C. M. (2013). RABL-regulated pathways: a new tale in sperm function. Asian J. Androl. 15, 87.
| RABL-regulated pathways: a new tale in sperm function.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXktFWrug%3D%3D&md5=38c449797e591bcfe5e688920531ae88CAS |
Blencowe, B. J. (2000). Exonic splicing enhancers: mechanism of action, diversity and role in human genetic diseases. Trends Biochem. Sci. 25, 106–110.
| Exonic splicing enhancers: mechanism of action, diversity and role in human genetic diseases.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXhsFOisr0%3D&md5=af11ced3cb72b279b7153bbfbc9dcad5CAS |
Caminsky, N., Mucaki, E. J., and Rogan, P. K. (2014). Interpretation of mRNA splicing mutations in genetic disease: review of the literature and guidelines for information-theoretical analysis. F1000Res 3, 282.
Cartegni, L., Chew, S. L., and Krainer, A. R. (2002). Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat. Rev. Genet. 3, 285–298.
| Listening to silence and understanding nonsense: exonic mutations that affect splicing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XjtFOrtrc%3D&md5=c1c0438db8cbf35ce1da4cc9cf5ff9faCAS |
Cartegni, L., Wang, J., Zhu, Z., Zhang, M. Q., and Krainer, A. R. (2003). ESEfinder: a web resource to identify exonic splicing enhancers. Nucleic Acids Res. 31, 3568–3571.
| ESEfinder: a web resource to identify exonic splicing enhancers.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXltVWisLw%3D&md5=e465794a2cb8d67908ad50392ef959beCAS |
Cassinello, J., Abaigar, T., Gomendio, M., and Roldan, E. R. (1998). Characteristics of the semen of three endangered species of gazelles (Gazella dama mhorr, G. dorcas neglecta and G. cuvieri). J. Reprod. Fertil. 113, 35–45.
| Characteristics of the semen of three endangered species of gazelles (Gazella dama mhorr, G. dorcas neglecta and G. cuvieri).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXlsVSgtbw%3D&md5=362f05148c58103edc8afec296920962CAS |
Cochran, S. D., Cole, J. B., Null, D. J., and Hansen, P. J. (2013). Discovery of single nucleotide polymorphisms in candidate genes associated with fertility and production traits in Holstein cattle. BMC Genet. 14, 49.
| Discovery of single nucleotide polymorphisms in candidate genes associated with fertility and production traits in Holstein cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhtVGqtbvM&md5=115276ade2711d7cf698a782816de251CAS |
Fairbrother, W. G., Yeh, R. F., Sharp, P. A., and Burge, C. B. (2002). Predictive identification of exonic splicing enhancers in human genes. Science 297, 1007–1013.
| Predictive identification of exonic splicing enhancers in human genes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xmt1Ckt74%3D&md5=90c47131eb1dcf9bedca737534e04e16CAS |
Gabut, M., Miné, M., Marsac, C., Brivet, M., Tazi, J., and Soret, J. (2005). The SR protein SC35 is responsible for aberrant splicing of the E1α pyruvate dehydrogenase mRNA in a case of mental retardation with lactic acidosis. Mol. Cell. Biol. 25, 3286–3294.
| The SR protein SC35 is responsible for aberrant splicing of the E1α pyruvate dehydrogenase mRNA in a case of mental retardation with lactic acidosis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXjtlSjs7s%3D&md5=5be36984bc9a161a7bdd811c7722bcf7CAS |
Gaffney, E. A., Gadêlha, H., Smith, D. J., Blake, J. R., and Kirkman-Brown, J. C. (2011). Mammalian sperm motility: observation and theory. Annu. Rev. Fluid Mech. 43, 501–528.
| Mammalian sperm motility: observation and theory.Crossref | GoogleScholarGoogle Scholar |
Gao, Q., Ju, Z., Zhang, Y., Huang, J., Zhang, X., Qi, C., Li, J., Zhong, J., Li, G., and Wang, C. (2014). Association of TNP2 gene polymorphisms of the bta-miR-154 target site with the semen quality traits of Chinese Holstein bulls. PLoS One 9, e84355.
| Association of TNP2 gene polymorphisms of the bta-miR-154 target site with the semen quality traits of Chinese Holstein bulls.Crossref | GoogleScholarGoogle Scholar |
Garcia-Blanco, M. A. (2006). Alternative splicing: therapeutic target and tool. Prog. Mol. Subcell. Biol. 44, 47–64.
| Alternative splicing: therapeutic target and tool.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhtFeitrnM&md5=d37e60c4368479833c9fcc5cfbeede7fCAS |
Garcia-Blanco, M. A., Baraniak, A. P., and Lasda, E. L. (2004). Alternative splicing in disease and therapy. Nat. Biotechnol. 22, 535–546.
| Alternative splicing in disease and therapy.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXjsF2jsLc%3D&md5=a84e90ab2b7b4523c2621d62aa04d908CAS |
Goddard, M. E., and Hayes, B. J. (2009). Mapping genes for complex traits in domestic animals and their use in breeding programmes. Nat. Rev. Genet. 10, 381–391.
| Mapping genes for complex traits in domestic animals and their use in breeding programmes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXmtVKrsLw%3D&md5=17679022f58a654c82491e90b1b21453CAS |
Guo, F., Yang, B., Ju, Z. H., Wang, X. G., Qi, C., Zhang, Y., Wang, C. F., Liu, H. D., Feng, M. Y., Chen, Y., Xu, Y. X., Zhong, J. F., and Huang, J. M. (2014). Alternative splicing, promoter methylation, and functional SNPs of sperm flagella 2 gene in testis and mature spermatozoa of Holstein bulls. Reproduction 147, 241–252.
| Alternative splicing, promoter methylation, and functional SNPs of sperm flagella 2 gene in testis and mature spermatozoa of Holstein bulls.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXisFartb8%3D&md5=e01b8d23e2aefeebda1c022f48569876CAS |
Hastings, M. L., and Krainer, A. R. (2001). Pre-mRNA splicing in the new millennium. Curr. Opin. Cell Biol. 13, 302–309.
| Pre-mRNA splicing in the new millennium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXktVaju7w%3D&md5=d9e863e96a9eb6efea2ea4f717dd1c06CAS |
Kramer, M., Huse, K., Menzel, U., Backhaus, O., Rosenstiel, P., Schreiber, S., Hampe, J., and Platzer, M. (2011). Constant splice-isoform ratios in human lymphoblastoid cells support the concept of a splico-stat. Genetics 187, 761–770.
| Constant splice-isoform ratios in human lymphoblastoid cells support the concept of a splico-stat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXmsVSgs7s%3D&md5=c0005a37eb8ef2e9992cdf9f50537daeCAS |
Liu, X., Ju, Z., Wang, L., Zhang, Y., Huang, J., Li, Q., Li, J., Zhong, J., An, L., and Wang, C. (2011). Six novel single-nucleotide polymorphisms in SPAG11 gene and their association with sperm quality traits in Chinese Holstein bulls. Anim. Reprod. Sci. 129, 14–21.
| Six novel single-nucleotide polymorphisms in SPAG11 gene and their association with sperm quality traits in Chinese Holstein bulls.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhsFOgu7%2FO&md5=6ce687fd4b155b741cd9c10381304be2CAS |
Lo, J. C., Jamsai, D., O’Connor, A. E., Borg, C., Clark, B. J., Whisstock, J. C., Field, M. C., Adams, V., Ishikawa, T., Aitken, R. J., Whittle, B., Goodnow, C. C., Ormandy, C. J., and O’Bryan, M. K. (2012). RAB-like 2 has an essential role in male fertility, sperm intra-flagellar transport, and tail assembly. PLoS Genet. 8, e1002969.
| RAB-like 2 has an essential role in male fertility, sperm intra-flagellar transport, and tail assembly.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhsFalu73K&md5=fb18198ee082bfa47d84b5a03fef9819CAS |
Long, J. C., and Caceres, J. F. (2009). The SR protein family of splicing factors: master regulators of gene expression. Biochem. J. 417, 15–27.
| The SR protein family of splicing factors: master regulators of gene expression.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsV2lt7%2FM&md5=e7a4dba4860620294f496c14ae996317CAS |
Ochs, M. J., Sorg, B. L., Pufahl, L., Grez, M., Suess, B., and Steinhilber, D. (2012). Post-transcriptional regulation of 5-lipoxygenase mRNA expression via alternative splicing and nonsense-mediated mRNA decay. PLoS One 7, e31363.
| Post-transcriptional regulation of 5-lipoxygenase mRNA expression via alternative splicing and nonsense-mediated mRNA decay.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjsVaqtb8%3D&md5=b8c71d8d2b0f055ed52fdce9a8c64047CAS |
Palhais, B., Præstegaard, V. S., Sabaratnam, R., Doktor, T. K., Lutz, S., Burda, P., Suormala, T., Baumgartner, M., Fowler, B., Bruun, G. H., Andersen, H. S., Kožich, V., and Andresen, B. S. (2015). Splice-shifting oligonucleotide (SSO) mediated blocking of an exonic splicing enhancer (ESE) created by the prevalent c.903+469T>C MTRR mutation corrects splicing and restores enzyme activity in patient cells. Nucleic Acids Res. 43, 4627–4639.
| Splice-shifting oligonucleotide (SSO) mediated blocking of an exonic splicing enhancer (ESE) created by the prevalent c.903+469T>C MTRR mutation corrects splicing and restores enzyme activity in patient cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhsFaisb%2FL&md5=56541e577bf82ec776e969997b0cd4b7CAS |
Qin, H., Wang, Z., Diener, D., and Rosenbaum, J. (2007). Intraflagellar transport protein 27 is a small G protein involved in cell-cycle control. Curr. Biol. 17, 193–202.
| Intraflagellar transport protein 27 is a small G protein involved in cell-cycle control.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtlOntL0%3D&md5=918bef86ee91c40c0cc80ef0d1d7485aCAS |
Stamm, S., Ben-Ari, S., Rafalska, I., Tang, Y., Zhang, Z., Toiber, D., Thanaraj, T. A., and Soreq, H. (2005). Function of alternative splicing. Gene 344, 1–20.
| Function of alternative splicing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXkvVSgtQ%3D%3D&md5=fda5ad426b6c169feeefe44efda523e2CAS |
Tazi, J., Bakkour, N., and Stamm, S. (2009). Alternative splicing and disease. Biochim. Biophys. Acta 1792, 14–26.
| Alternative splicing and disease.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsFClsrbP&md5=9672efceaf25b6b2d0bfaf89fdcd4efaCAS |
Turner, R. M. (2006). Moving to the beat: a review of mammalian sperm motility regulation. Reprod. Fertil. Dev. 18, 25–38.
| Moving to the beat: a review of mammalian sperm motility regulation.Crossref | GoogleScholarGoogle Scholar |
Venables, J. P. (2004). Aberrant and alternative splicing in cancer. Cancer Res. 64, 7647–7654.
| Aberrant and alternative splicing in cancer.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXptFemtb8%3D&md5=1608883c846023ca52295ec7d9a8715dCAS |
Wang, X., Zhong, J., Gao, Y., Ju, Z., and Huang, J. (2014). A SNP in intron 8 of CD46 causes a novel transcript associated with mastitis in Holsteins. BMC Genomics 15, 630.
| A SNP in intron 8 of CD46 causes a novel transcript associated with mastitis in Holsteins.Crossref | GoogleScholarGoogle Scholar |
Weigel, K. A. (2006). Prospects for improving reproductive performance through genetic selection. Anim. Reprod. Sci. 96, 323–330.
| Prospects for improving reproductive performance through genetic selection.Crossref | GoogleScholarGoogle Scholar |
