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. 1997 Jan 15;321(Pt 2):325–332. doi: 10.1042/bj3210325

Isolation and characterization of the androgen-dependent mouse cysteine-rich secretory protein-1 (CRISP-1) gene.

U Schwidetzky 1, W D Schleuning 1, B Haendler 1
PMCID: PMC1218072  PMID: 9020862

Abstract

In mice, cysteine-rich secretory protein-1 (CRISP-1) is mainly found in the epididymis and also, to a lesser extent, in the salivary gland of males, where androgens control its expression. We have now isolated and characterized overlapping phage clones covering the entire length of the CRISP-1 gene. DNA sequencing revealed that the gene is organized into eight exons, ranging between 55 and 748 bp in size, and seven introns. All exon-intron junctions conformed to the GT/AG rule established for eukaryotic genes. The intron length, as determined by PCR, varied between 1.05 and 4.0 kb so that the CRISP-1 gene spans over 20 kb of the mouse genome. The transcription-initiation site was determined by primer extension and localized at the expected distance downstream of a consensus TATA box. Approximately 3.7 kb of the CRISP-1 promoter region were isolated and sequenced, and several stretches fitting the androgen-responsive element consensus were found. Those that most resembled the consensus were analysed by electrophoretic mobility-shift assay and found to form specific complexes with the liganded androgen receptor in vitro, but with different affinities. Putative binding elements for the transcription factors Oct, GATA, PEA3, CF1. AP-1 and AP-3 were also found in the promoter region.

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Selected References

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  1. Adler A. J., Scheller A., Hoffman Y., Robins D. M. Multiple components of a complex androgen-dependent enhancer. Mol Endocrinol. 1991 Nov;5(11):1587–1596. doi: 10.1210/mend-5-11-1587. [DOI] [PubMed] [Google Scholar]
  2. Adler A. J., Scheller A., Robins D. M. The stringency and magnitude of androgen-specific gene activation are combinatorial functions of receptor and nonreceptor binding site sequences. Mol Cell Biol. 1993 Oct;13(10):6326–6335. doi: 10.1128/mcb.13.10.6326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ariel M., Cedar H., McCarrey J. Developmental changes in methylation of spermatogenesis-specific genes include reprogramming in the epididymis. Nat Genet. 1994 May;7(1):59–63. doi: 10.1038/ng0594-59. [DOI] [PubMed] [Google Scholar]
  4. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Brooks D. E. Androgen-regulated epididymal secretory proteins associated with post-testicular sperm development. Ann N Y Acad Sci. 1987;513:179–194. doi: 10.1111/j.1749-6632.1987.tb25008.x. [DOI] [PubMed] [Google Scholar]
  7. Brooks D. E., Means A. R., Wright E. J., Singh S. P., Tiver K. K. Molecular cloning of the cDNA for androgen-dependent sperm-coating glycoproteins secreted by the rat epididymis. Eur J Biochem. 1986 Nov 17;161(1):13–18. doi: 10.1111/j.1432-1033.1986.tb10118.x. [DOI] [PubMed] [Google Scholar]
  8. Brooks D. E., Tiver K. Localization of epididymal secretory proteins on rat spermatozoa. J Reprod Fertil. 1983 Nov;69(2):651–657. doi: 10.1530/jrf.0.0690651. [DOI] [PubMed] [Google Scholar]
  9. Brüggemeier U., Kalff M., Franke S., Scheidereit C., Beato M. Ubiquitous transcription factor OTF-1 mediates induction of the MMTV promoter through synergistic interaction with hormone receptors. Cell. 1991 Feb 8;64(3):565–572. doi: 10.1016/0092-8674(91)90240-y. [DOI] [PubMed] [Google Scholar]
  10. Bucher P., Trifonov E. N. Compilation and analysis of eukaryotic POL II promoter sequences. Nucleic Acids Res. 1986 Dec 22;14(24):10009–10026. doi: 10.1093/nar/14.24.10009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cameo M. S., Blaquier J. A. Androgen-controlled specific proteins in rat epididymis. J Endocrinol. 1976 Apr;69(1):47–55. doi: 10.1677/joe.0.0690047. [DOI] [PubMed] [Google Scholar]
  12. Cato A. C., Skroch P., Weinmann J., Butkeraitis P., Ponta H. DNA sequences outside the receptor-binding sites differently modulate the responsiveness of the mouse mammary tumour virus promoter to various steroid hormones. EMBO J. 1988 May;7(5):1403–1410. doi: 10.1002/j.1460-2075.1988.tb02957.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cooper T. G. In defense of a function for the human epididymis. Fertil Steril. 1990 Dec;54(6):965–975. doi: 10.1016/s0015-0282(16)53988-0. [DOI] [PubMed] [Google Scholar]
  14. Corden J., Wasylyk B., Buchwalder A., Sassone-Corsi P., Kedinger C., Chambon P. Promoter sequences of eukaryotic protein-coding genes. Science. 1980 Sep 19;209(4463):1406–1414. doi: 10.1126/science.6251548. [DOI] [PubMed] [Google Scholar]
  15. Culig Z., Hobisch A., Cronauer M. V., Cato A. C., Hittmair A., Radmayr C., Eberle J., Bartsch G., Klocker H. Mutant androgen receptor detected in an advanced-stage prostatic carcinoma is activated by adrenal androgens and progesterone. Mol Endocrinol. 1993 Dec;7(12):1541–1550. doi: 10.1210/mend.7.12.8145761. [DOI] [PubMed] [Google Scholar]
  16. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Eberspaecher U., Roosterman D., Krätzschmar J., Haendler B., Habenicht U. F., Becker A., Quensel C., Petri T., Schleuning W. D., Donner P. Mouse androgen-dependent epididymal glycoprotein CRISP-1 (DE/AEG): isolation, biochemical characterization, and expression in recombinant form. Mol Reprod Dev. 1995 Oct;42(2):157–172. doi: 10.1002/mrd.1080420205. [DOI] [PubMed] [Google Scholar]
  18. Faisst S., Meyer S. Compilation of vertebrate-encoded transcription factors. Nucleic Acids Res. 1992 Jan 11;20(1):3–26. doi: 10.1093/nar/20.1.3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Foster J. A., Gerton G. L. Autoantigen 1 of the guinea pig sperm acrosome is the homologue of mouse Tpx-1 and human TPX1 and is a member of the cysteine-rich secretory protein (CRISP) family. Mol Reprod Dev. 1996 Jun;44(2):221–229. doi: 10.1002/(SICI)1098-2795(199606)44:2<221::AID-MRD11>3.0.CO;2-5. [DOI] [PubMed] [Google Scholar]
  20. Fournier-Delpech S., Courot M., Dubois M. P. Decreased fertility and motility of spermatozoa from rats immunized with a prealbumin epididymal-specific glycoprotein. J Androl. 1985 Jul-Aug;6(4):246–250. doi: 10.1002/j.1939-4640.1985.tb00841.x. [DOI] [PubMed] [Google Scholar]
  21. Ghyselinck N. B., Dufaure I., Lareyre J. J., Rigaudière N., Mattéi M. G., Dufaure J. P. Structural organization and regulation of the gene for the androgen-dependent glutathione peroxidase-like protein specific to the mouse epididymis. Mol Endocrinol. 1993 Feb;7(2):258–272. doi: 10.1210/mend.7.2.8469239. [DOI] [PubMed] [Google Scholar]
  22. Haendler B., Krätzschmar J., Theuring F., Schleuning W. D. Transcripts for cysteine-rich secretory protein-1 (CRISP-1; DE/AEG) and the novel related CRISP-3 are expressed under androgen control in the mouse salivary gland. Endocrinology. 1993 Jul;133(1):192–198. doi: 10.1210/endo.133.1.8319566. [DOI] [PubMed] [Google Scholar]
  23. Hardy D. M., Huang T. T., Jr, Driscoll W. J., Tung K. K., Wild G. C. Purification and characterization of the primary acrosomal autoantigen of guinea pig epididymal spermatozoa. Biol Reprod. 1988 Mar;38(2):423–437. doi: 10.1095/biolreprod38.2.423. [DOI] [PubMed] [Google Scholar]
  24. Hayashi M., Fujimoto S., Takano H., Ushiki T., Abe K., Ishikura H., Yoshida M. C., Kirchhoff C., Ishibashi T., Kasahara M. Characterization of a human glycoprotein with a potential role in sperm-egg fusion: cDNA cloning, immunohistochemical localization, and chromosomal assignment of the gene (AEGL1). Genomics. 1996 Mar 15;32(3):367–374. doi: 10.1006/geno.1996.0131. [DOI] [PubMed] [Google Scholar]
  25. Jänne O. A., Palvimo J. J., Kallio P., Mehto M. Androgen receptor and mechanism of androgen action. Ann Med. 1993 Feb;25(1):83–89. doi: 10.3109/07853899309147863. [DOI] [PubMed] [Google Scholar]
  26. Kasahara M., Gutknecht J., Brew K., Spurr N., Goodfellow P. N. Cloning and mapping of a testis-specific gene with sequence similarity to a sperm-coating glycoprotein gene. Genomics. 1989 Oct;5(3):527–534. doi: 10.1016/0888-7543(89)90019-0. [DOI] [PubMed] [Google Scholar]
  27. Kasahara M., Hayashi M., Yoshida M. C., Nadeau J. H., Fujimoto S., Ishibashi T. Mapping of acidic epididymal glycoprotein (Aeg) genes to mouse chromosome 17. Mamm Genome. 1995 Jan;6(1):52–54. doi: 10.1007/BF00350895. [DOI] [PubMed] [Google Scholar]
  28. Keller E. B., Noon W. A. Intron splicing: a conserved internal signal in introns of animal pre-mRNAs. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7417–7420. doi: 10.1073/pnas.81.23.7417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kjeldsen L., Cowland J. B., Johnsen A. H., Borregaard N. SGP28, a novel matrix glycoprotein in specific granules of human neutrophils with similarity to a human testis-specific gene product and a rodent sperm-coating glycoprotein. FEBS Lett. 1996 Feb 19;380(3):246–250. doi: 10.1016/0014-5793(96)00030-0. [DOI] [PubMed] [Google Scholar]
  30. Krätzschmar J., Haendler B., Eberspaecher U., Roosterman D., Donner P., Schleuning W. D. The human cysteine-rich secretory protein (CRISP) family. Primary structure and tissue distribution of CRISP-1, CRISP-2 and CRISP-3. Eur J Biochem. 1996 Mar 15;236(3):827–836. doi: 10.1111/j.1432-1033.1996.t01-1-00827.x. [DOI] [PubMed] [Google Scholar]
  31. Lancillotti F., Lopez M. C., Arias P., Alonso C. Z-DNA in transcriptionally active chromosomes. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1560–1564. doi: 10.1073/pnas.84.6.1560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mizuki N., Kasahara M. Mouse submandibular glands express an androgen-regulated transcript encoding an acidic epididymal glycoprotein-like molecule. Mol Cell Endocrinol. 1992 Nov;89(1-2):25–32. doi: 10.1016/0303-7207(92)90207-m. [DOI] [PubMed] [Google Scholar]
  33. Mochca-Morales J., Martin B. M., Possani L. D. Isolation and characterization of helothermine, a novel toxin from Heloderma horridum horridum (Mexican beaded lizard) venom. Toxicon. 1990;28(3):299–309. doi: 10.1016/0041-0101(90)90065-f. [DOI] [PubMed] [Google Scholar]
  34. Montgomery J. C., Venta P. J., Eddy R. L., Fukushima Y. S., Shows T. B., Tashian R. E. Characterization of the human gene for a newly discovered carbonic anhydrase, CA VII, and its localization to chromosome 16. Genomics. 1991 Dec;11(4):835–848. doi: 10.1016/0888-7543(91)90006-z. [DOI] [PubMed] [Google Scholar]
  35. Morrissette J., Krätzschmar J., Haendler B., el-Hayek R., Mochca-Morales J., Martin B. M., Patel J. R., Moss R. L., Schleuning W. D., Coronado R. Primary structure and properties of helothermine, a peptide toxin that blocks ryanodine receptors. Biophys J. 1995 Jun;68(6):2280–2288. doi: 10.1016/S0006-3495(95)80410-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Patthy L. Intron-dependent evolution: preferred types of exons and introns. FEBS Lett. 1987 Apr 6;214(1):1–7. doi: 10.1016/0014-5793(87)80002-9. [DOI] [PubMed] [Google Scholar]
  37. Perez Martinez S., Conesa D., Cuasnicú P. S. Potential contraceptive use of epididymal proteins: evidence for the participation of specific antibodies against rat epididymal protein DE in male and female fertility inhibition. J Reprod Immunol. 1995 May;29(1):31–45. doi: 10.1016/0165-0378(95)00927-d. [DOI] [PubMed] [Google Scholar]
  38. Pfisterer P., König H., Hess J., Lipowsky G., Haendler B., Schleuning W. D., Wirth T. CRISP-3, a protein with homology to plant defense proteins, is expressed in mouse B cells under the control of Oct2. Mol Cell Biol. 1996 Nov;16(11):6160–6168. doi: 10.1128/mcb.16.11.6160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rankin T. L., Tsuruta K. J., Holland M. K., Griswold M. D., Orgebin-Crist M. C. Isolation, immunolocalization, and sperm-association of three proteins of 18, 25, and 29 kilodaltons secreted by the mouse epididymis. Biol Reprod. 1992 May;46(5):747–766. doi: 10.1095/biolreprod46.5.747. [DOI] [PubMed] [Google Scholar]
  40. Rich A., Nordheim A., Wang A. H. The chemistry and biology of left-handed Z-DNA. Annu Rev Biochem. 1984;53:791–846. doi: 10.1146/annurev.bi.53.070184.004043. [DOI] [PubMed] [Google Scholar]
  41. Rigden J., Coutts R. Pathogenesis-related proteins in plants. Trends Genet. 1988 Apr;4(4):87–89. doi: 10.1016/0168-9525(88)90091-1. [DOI] [PubMed] [Google Scholar]
  42. Robaire B., Viger R. S. Regulation of epididymal epithelial cell functions. Biol Reprod. 1995 Feb;52(2):226–236. doi: 10.1095/biolreprod52.2.226. [DOI] [PubMed] [Google Scholar]
  43. Roche P. J., Hoare S. A., Parker M. G. A consensus DNA-binding site for the androgen receptor. Mol Endocrinol. 1992 Dec;6(12):2229–2235. doi: 10.1210/mend.6.12.1491700. [DOI] [PubMed] [Google Scholar]
  44. Rochwerger L., Cohen D. J., Cuasnicú P. S. Mammalian sperm-egg fusion: the rat egg has complementary sites for a sperm protein that mediates gamete fusion. Dev Biol. 1992 Sep;153(1):83–90. doi: 10.1016/0012-1606(92)90093-v. [DOI] [PubMed] [Google Scholar]
  45. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Scarlett C. O., Robins D. M. In vivo footprinting of an androgen-dependent enhancer reveals an accessory element integral to hormonal response. Mol Endocrinol. 1995 Apr;9(4):413–423. doi: 10.1210/mend.9.4.7659085. [DOI] [PubMed] [Google Scholar]
  47. Schwidetzky U., Haendler B., Schleuning W. D. Isolation and characterization of the androgen-dependent mouse cysteine-rich secretory protein-3 (CRISP-3) gene. Biochem J. 1995 Aug 1;309(Pt 3):831–836. doi: 10.1042/bj3090831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Traut T. W. Do exons code for structural or functional units in proteins? Proc Natl Acad Sci U S A. 1988 May;85(9):2944–2948. doi: 10.1073/pnas.85.9.2944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Triggs-Raine B. L., Benoit G., Salo T. J., Trasler J. M., Gravel R. A. Characterization of the murine beta-hexosaminidase (HEXB) gene. Biochim Biophys Acta. 1994 Oct 21;1227(1-2):79–86. doi: 10.1016/0925-4439(94)90110-4. [DOI] [PubMed] [Google Scholar]
  50. Vierula M. E., Rankin T. L., Orgebin-Crist M. C. Electron microscopic immunolocalization of the 18 and 29 kilodalton secretory proteins in the mouse epididymis: evidence for differential uptake by clear cells. Microsc Res Tech. 1995 Jan 1;30(1):24–36. doi: 10.1002/jemt.1070300103. [DOI] [PubMed] [Google Scholar]
  51. Vreeburg J. T., Holland M. K., Orgebin-Crist M. C. Binding of epididymal proteins to rat spermatozoa in vivo. Biol Reprod. 1992 Oct;47(4):588–597. doi: 10.1095/biolreprod47.4.588. [DOI] [PubMed] [Google Scholar]
  52. Xie Y. B., Sui Y. P., Shan L. X., Palvimo J. J., Phillips D. M., Jänne O. A. Expression of androgen receptor in insect cells. Purification of the receptor and renaturation of its steroid- and DNA-binding functions. J Biol Chem. 1992 Mar 5;267(7):4939–4948. [PubMed] [Google Scholar]
  53. Xin J. H., Cowie A., Lachance P., Hassell J. A. Molecular cloning and characterization of PEA3, a new member of the Ets oncogene family that is differentially expressed in mouse embryonic cells. Genes Dev. 1992 Mar;6(3):481–496. doi: 10.1101/gad.6.3.481. [DOI] [PubMed] [Google Scholar]
  54. Xu L. C., Thali M., Schaffner W. Upstream box/TATA box order is the major determinant of the direction of transcription. Nucleic Acids Res. 1991 Dec 25;19(24):6699–6704. doi: 10.1093/nar/19.24.6699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zeitlin S., Efstratiadis A. In vivo splicing products of the rabbit beta-globin pre-mRNA. Cell. 1984 Dec;39(3 Pt 2):589–602. doi: 10.1016/0092-8674(84)90466-5. [DOI] [PubMed] [Google Scholar]

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