Abstract
Tpx-1 is a testis-specific gene that maps on mouse Chromosome (Chr) 17. The deduced TPX-1 protein shows 55% amino acid sequence similarity to acidic epididymal glycoprotein (AEG), assumed to be involved in sperm maturation. In the present study, we determined the genomic structure of the mouse Tpx-1 gene and the cellular localization of its transcripts. The gene was found to contain ten exons, with an unusually large intron (∼17.0 kilobase pairs) between exons 8 and 9. In situ hybridization of testicular sections showed that Tpx-1 is transcribed abundantly by haploid male germ cells. A computer search of protein databases revealed that deduced TPX-1/AEG proteins have significant sequence similarity (∼30%) to two non-mammalian proteins: “pathogenesis-related” proteins 1 of tobaccos, and venom sac proteins of whiteface hornets, known as Dol m V. Amino acid residues encoded by exon 10 of the Tpx-1 gene and most of those encoded by exon 9 were absent in the non-mammalian proteins. This result suggests that the ancestor of Tpx-1 acquired exons 9 and 10 after its divergence from the ancestors of the plant and insect proteins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bedford, J.M.: Changes in the electrophoretic properties of rabbit spermatozoa during passage through the epididymis. Nature 200: 1178–1180, 1963.
Bedford, J.M.: Maturation, transport, and fate of spermatozoa in the epididymis. In D.W. Hamilton and R.O. Greep (eds), Handbook of Physiology, Vol. 5, pp. 303–317, American Physiological Society, Washington DC, 1975.
Bellvé, A.R.: The molecular biology of mammalian spermatogenesis. Oxford Rev Reprod Biol 1: 159–261, 1979.
Bowles, D.J.: Defense-related proteins in higher plants. Annu Rev Biochem 59: 873–907, 1990.
Breathnach, R., Benoist, C., O'Hare, K., Gannon, F., and Chambon, P.: Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci USA 75: 4853–4857, 1978.
Brooks, D.E.: Purification of rat epididymal proteins ‘D’ and ‘E’, demonstration of shared immunological determinants, and identification of regional synthesis and secretion. Int J Androl 5: 513–524, 1982.
Brooks, D.E. and Tiver, K.: Localization of epididymal secretory proteins on rat spermatozoa. J Reprod Fertil 69: 651–657, 1983.
Brooks, D.E., Means, A.R., Wright, E.J., Singh, S.P., and Tiver, K.K.: Molecular cloning of the cDNA for androgen-dependent sperm-coating glycoprotein secreted by the rat epididymis. Eur J Biochem 161: 13–18, 1986.
Brutlag, D.L., Dautricourt, J.-P., Maulik, S., and Relph, J.: Improved sensitivity of biological sequence database searches. CABIOS 6, 237–245, 1990.
Cameo, M.S. and Blaquier, J.A.: Androgen-controlled specific proteins in rat epididymis. J Endocrinol 69: 47–55, 1976.
Charest, N.J., Joseph, D.R., Wilson, E.M., and French, F.S.: Molecular cloning of complementary deoxyribonucleic acid for an androgen-regulated epididymal protein: sequence homology with metalloproteins. Mol Endocrinol 2: 999–1004, 1988.
Clermont, Y. and Tang, X.M.: Glycoprotein synthesis in the Golgi apparatus of spermatids during spermiogenesis of the rat. Anat Rec 213: 33–43, 1985.
Cornelissen, B.J.C., Horowitz, J., van Kan, J.A.L., Goldberg, R.B., and Bol, J.F.: Structures of tobacco genes encoding pathogenesis-related proteins from the PR-1 group. Nucleic Acids Res 15: 6799–6811, 1987.
Craik, C.S., Rutter, W.J., and Fletterick, R.: Splice junctions: association with variation in protein structure. Science 220: 1125–1129, 1983.
Domenjoud, L., Nussbaum, G., Adham, I.M., Greeske, G., and Engel, W.: Genomic sequence of human protamines whose genes, PRM1 and PRM2, are clustered. Genomics 8: 127–133, 1990.
Doolittle, R.F.: Searching through sequence databases. Methods Enzymol 183: 99–110, 1990.
Erickson, R.P.: Post-meiotic gene expression. Trends Genet 6: 264–269, 1990.
Evans, G.A. and Wahl, G.M.: Cosmid vectors for genomic walking and rapid restriction mapping. Methods Enzymol 152: 604–610, 1987.
Fang, K.S.Y., Vitale, M., Fehlner, P., and King, T.P.: cDNA cloning and primary structure of a white-face hornet venom allergen, antigen 5. Proc Natl Acad Sci USA 85: 895–899, 1988.
Faye, J.C., Duguet, L., Mazzuca, M., and Bayard, F.: Purification, radioimmunoassay, and immunohistochemical localization of a glycoprotein produced by the rat epididymis. Biol Reprod 23: 423–432, 1980.
Jenne, D.E., Zimmer, M., Garcia-Sanz, J.A., Tschopp, J., and Lichter, P.: Genomic organization and subchromosomal in situ localization of the murine granzyme F, a serine protease expressed in CD8+ T cells. J. Immunol 147: 1045–1052, 1991.
Johnson, P.A., Peschon, J.J., Yelick, P.C. Palmiter, R.D., and Hecht, N.B.: Sequence homologies in the mouse protamine 1 and 2 genes. Biochim Biophys Acta 950: 45–53, 1988.
Jones, R., Brown, C.R., von Glos, K.I., and Parker, M.G.: Hormonal regulation of protein synthesis in the rat epididymis. Biochem J 188: 667–676, 1980.
Kadonaga, J.T., Jones, K.A., and Tjian, R.: Promoter-specific activation of RNA polymerase II transcription by Sp1. Trends Biochem Sci 11: 20–23, 1986.
Kasahara, M., Figueroa, F., and Klein, J.: Random cloning of genes from mouse chromosome 17. Proc Natl Acad Sci USA 84: 3325–3328, 1987.
Kasahara, M., Passmore, H.C., and Klein, J.: A testis-specific gene Tpx-1 maps between Pgk-2 and Mep-1 on mouse chromosome 17. Immunogenetics 29: 61–63, 1989a.
Kasahara, M., Gutknecht, J., Brew, K., Spurr, N., and Goodfellow, P.N.: Cloning and mapping of a testis-specific gene with sequence similarity to a sperm-coating glycoprotein gene. Genomics 5: 527–534, 1989b.
Kashiwabara, S., Baba, T., Takada, M., Watanabe, K., Yano, Y., and Arai, Y.: Primary structure of mouse proacrosin deduced from the cDNA sequence and its gene expression during spermatogenesis. J Biochem 108: 785–791, 1990a.
Kashiwabara, S., Arai, Y., Kodaira, K., and Baba, T.: Acrosin biosynthesis in meiotic and postmeiotic spermatogenic cells. Biochem Biophys Res Commun 173: 240–245, 1990b.
Klemm, U., Maier, W.-M., Tsaousidou, S., Adham, I.M., Willison, K., and Engel, W.: Mouse preproacrosin: cDNA sequence, primary structure and postmeiotic expression in spermatogenesis. Differentiation 42: 160–166, 1990.
Kremling, H., Keime, S., Wilhelm, K., Adham, I.M., Hameister, H., and Engel, W.: Mouse proacrosin gene: nucleotide sequence, diploid expression, and chromosomal localization. Genomics 11: 828–834, 1991.
Land, H., Grez, M., Hauser, H., Lindenmaier, W., and Schütz, G.: 5′-Terminal sequences of eukaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res 9: 2251–2266, 1981.
Lea, O.A. and French, F.S.: Characterization of an acidic glyco-protein secreted by principal cells of the rat epididymis. Biochim Biophys Acta 668: 370–376, 1981.
Lee, K.A.W., Hai, T.Y., SivaRaman, L., Thimmappaya, B., Hurst, H.C., Jones, N.C., and Green, M.R.: A cellular transcription factor ATF activates transcription of multiple E1A-inducible adenovirus early promoters. Proc Natl Acad Sci USA 84: 8355–8359, 1987.
McCarrey, J.R.: Nucleotide sequence of the promoter region of a tissue-specific human retroposon: comparison with its housekeeping progenitor. Gene 61: 291–298, 1987.
Melton, D.A., Krieg, P.A., Rebagliati, M.R., Maniatis, T., Zinn, K., and Green, M.R.: Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12: 7035–7056, 1984.
Montminy, M.R., and Bilezikjian, L.M.: Binding of a nuclear protein to the cyclic AMP response element of the somatostatin gene. Nature 328: 175–178, 1987.
Mount, S.M.: A catalogue of splice junction sequences. Nucleic Acids Res 10: 459–472, 1982.
Oakberg, E.F.: A description of spermiogenesis in the mouse and its use in analysis of the cycle of the seminiferous epithelium and germ cell renewal. Am J Anat 99: 391–413, 1956a.
Oakberg, E.F.: Duration of spermatogenesis in the mouse and timing of stages of the cycle of the seminiferous epithelium. Am J Anat 99: 507–516, 1956b.
Ohshima, M., Matsuoka, M., Yamamoto, N., Tanaka, Y., Kano-Murakami, Y., Ozeki, Y., Kato, A., Harada, N., and Ohashi, Y.: Nucleotide sequence of the PR-1 gene of Nicotiana tabacum. FEBS Lett 225: 243–246, 1987.
Orgebin-Crist, M.-C., Danzo, B.J., and Davies, J.: Endocrine control of the development and maintenance of sperm fertilizing ability in the epididymis. In D.W. Hamilton and R.O. Greep (eds.); Handbook of Physiology, Vol. 5, pp. 319–338, American Physiological Society, Washington DC, 1975.
Pelham, H.R.B.: Activation of heat-shock genes in eukaryotes. Trends Genet 1: 31–35, 1985.
Pfitzner, U.M. and Goodman, H.M.: Isolation and characterization of cDNA clones encoding pathogenesis-related proteins from tobacco mosaic virus-infected tobacco plants. Nucleic Acids Res 15: 4449–4465, 1987.
Sanger, F., Nicklen, S., and Coulson, A.R.: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977.
Tartof, K.D. and Hobbs, C.A.: New cloning vectors and techniques for easy and rapid restriction mapping. Gene 67: 169–182, 1988.
Willison, K. and Ashworth, A.: Mammalian spermatogenic gene expression. Trends Genet 3: 351–355, 1987.
Wong, P.Y.D. and Tsang, A.Y.F.: Studies on the binding of a 32K rat epididymal protein to rat epididymal spermatozoa. Biol Reprod 27: 1239–1246, 1982.
Yanagimachi, R., Noda, Y.D., Fujimoto, M., and Nicholson, G.L.: The distribution of negative charges on mammalian spermatozoa. Am J Anat 135: 497–520, 1972.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mizuki, N., Sarapata, D.E., Garcia-Sanz, J.A. et al. The mouse male germ cell-specific gene Tpx-1: molecular structure, mode of expression in spermatogenesis, and sequence similarity to two non-mammalian genes. Mammalian Genome 3, 274–280 (1992). https://doi.org/10.1007/BF00292155
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00292155