Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Feb 25;11(4):917–930. doi: 10.1093/nar/11.4.917

Isolation and characterisation of genes for androgen-responsive secretory proteins of rat seminal vesicles.

C McDonald, L Williams, P McTurk, F Fuller, E McIntosh, S Higgins
PMCID: PMC325767  PMID: 6298734

Abstract

Under the influence of testosterone, rat seminal vesicles synthesise large amounts of a tissue specific protein, S. Recombinant lambda clones have been isolated containing overlapping sequences covering a 27.5 kilo base region of the rat genome within which the gene for protein S is located. Recombinant plasmids bearing cDNA sequences for protein S were constructed in pBR328. One (pcS2) contains a 690 nucleotide insert and is probably full length. Detailed restriction maps of the S-gene are presented and the structure was confirmed by analysis of R-loops and heteroduplexes. The S-gene covers a 2 kbp region of the genome and consists of a 5' intron (490 bp) separating a leading exon (120 bp) containing the 5' untranslated region from a central exon (310 bp) containing most of the coding sequence and part of the 3' untranslated region. A larger intron (1100 bp) lies within the 3' untranslated region. The cloned gene is representative of the native gene but the S gene may be heterogeneous. Using pcS2, the hormonal control of S-specific mRNA was examined and a pronounced differential response to testosterone was observed.

Full text

PDF
917

Images in this article

920Image
on p.920
922Image
on p.922
925Image
on p.925
926Image
on p.926
927Image
on p.927

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrescia P., Guardiola J., Felsani A., Metafora S. Expression in male and genomic organization of the gene(s) coding for a major protein secreted by the rat seminal vesicle epithelium. Nucleic Acids Res. 1982 Feb 25;10(4):1159–1174. doi: 10.1093/nar/10.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
  3. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark A. J., Clissold P. M., Bishop J. O. Variation between mouse major urinary protein genes isolated from a single inbred line. Gene. 1982 Jun;18(3):221–230. doi: 10.1016/0378-1119(82)90159-7. [DOI] [PubMed] [Google Scholar]
  6. Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Durnam D. M., Perrin F., Gannon F., Palmiter R. D. Isolation and characterization of the mouse metallothionein-I gene. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6511–6515. doi: 10.1073/pnas.77.11.6511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Higgins S. J., Burchell J. M. Effects of testosterone on messenger ribonucleic acid and protein synthesis in rat seminal vesicle. Biochem J. 1978 Aug 15;174(2):543–551. doi: 10.1042/bj1740543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Higgins S. J., Burchell J. M., Mainwaring W. I. Testosterone control of nucleic acid content and proliferation of epithelium and stroma in rat seminal vesicles. Biochem J. 1976 Oct 15;160(1):43–48. doi: 10.1042/bj1600043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Higgins S. J., Burchell J. M., Parker M. G., Herries D. G. Effects of testosterone on sequence complexity of polyadenylated RNA from rat seminal vesicle. Eur J Biochem. 1978 Nov 15;91(2):327–334. doi: 10.1111/j.1432-1033.1978.tb12683.x. [DOI] [PubMed] [Google Scholar]
  12. Higgins S. J., Fuller F. M. Effects of testosterone on protein synthesis in rat seminal vesicles analysed by two-dimensional gel electrophoresis. Mol Cell Endocrinol. 1981 Oct;24(1):85–101. doi: 10.1016/0303-7207(81)90081-2. [DOI] [PubMed] [Google Scholar]
  13. Jeffreys A. J., Flavell R. A. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. doi: 10.1016/0092-8674(77)90119-2. [DOI] [PubMed] [Google Scholar]
  14. Kistler M. K., Taylor R. E., Jr, Kandala J. C., Kistler W. S. Isolation of recombinant plasmids containing structural gene sequences for rat seminal vesicle secretory proteins IV and V. Biochem Biophys Res Commun. 1981 Apr 30;99(4):1161–1166. doi: 10.1016/0006-291x(81)90740-3. [DOI] [PubMed] [Google Scholar]
  15. Land H., Grez M., Hauser H., Lindenmaier W., Schütz G. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res. 1981 May 25;9(10):2251–2266. doi: 10.1093/nar/9.10.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mansson P. E., Sugino A., Harris S. E. Use of a cloned double stranded cDNA coding for a major androgen dependent protein in rat seminal vesicle secretion: the effect of testosterone in gene expression. Nucleic Acids Res. 1981 Feb 25;9(4):935–946. doi: 10.1093/nar/9.4.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  18. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  19. Naora H., Deacon N. J. Relationship between the total size of exons and introns in protein-coding genes of higher eukaryotes. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6196–6200. doi: 10.1073/pnas.79.20.6196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Page M. J., Parker M. G. Effect of androgen on the transcription of rat prostatic binding protein genes. Mol Cell Endocrinol. 1982 Aug;27(3):343–355. doi: 10.1016/0303-7207(82)90099-5. [DOI] [PubMed] [Google Scholar]
  21. Pan Y. C., Silverberg A. B., Harris S. E., Li S. S. Complete amino acid sequence of a major secretory protein from rat seminal vesicle. Int J Pept Protein Res. 1980 Aug;16(2):143–146. doi: 10.1111/j.1399-3011.1980.tb02947.x. [DOI] [PubMed] [Google Scholar]
  22. Parker M. G., White R., Williams J. G. Cloning and characterization of androgen-dependent mRNA from rat ventral prostate. J Biol Chem. 1980 Jul 25;255(14):6996–7001. [PubMed] [Google Scholar]
  23. Parker M., Needham M., White R., Hurst H., Page M. Prostatic steroid binding protein: organisation of C1 and C2 genes. Nucleic Acids Res. 1982 Sep 11;10(17):5121–5132. doi: 10.1093/nar/10.17.5121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Prentki P., Karch F., Iida S., Meyer J. The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene. 1981 Sep;14(4):289–299. doi: 10.1016/0378-1119(81)90161-x. [DOI] [PubMed] [Google Scholar]
  25. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  26. Royal A., Garapin A., Cami B., Perrin F., Mandel J. L., LeMeur M., Brégégègre F., Gannon F., LePennec J. P., Chambon P. The ovalbumin gene region: common features in the organisation of three genes expressed in chicken oviduct under hormonal control. Nature. 1979 May 10;279(5709):125–132. doi: 10.1038/279125a0. [DOI] [PubMed] [Google Scholar]
  27. Sargent T. D., Wu J. R., Sala-Trepat J. M., Wallace R. B., Reyes A. A., Bonner J. The rat serum albumin gene: analysis of cloned sequences. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3256–3260. doi: 10.1073/pnas.76.7.3256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schafer M. P., Boyd C. D., Tolstoshev P., Crystal R. G. Structural organization of a 17 KB segment of the alpha 2 collagen gene: evaluation by R loop mapping. Nucleic Acids Res. 1980 May 24;8(10):2241–2253. doi: 10.1093/nar/8.10.2241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Soberon X., Covarrubias L., Bolivar F. Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. Gene. 1980 May;9(3-4):287–305. doi: 10.1016/0378-1119(90)90328-o. [DOI] [PubMed] [Google Scholar]
  30. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  31. Staden R. Sequence data handling by computer. Nucleic Acids Res. 1977 Nov;4(11):4037–4051. doi: 10.1093/nar/4.11.4037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Swaneck G. E., Nordstrom J. L., Kreuzaler F., Tsai M. J., O'Malley B. W. Effect of estrogen on gene expression in chicken oviduct: evidence for transcriptional control of ovalbumin gene. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1049–1053. doi: 10.1073/pnas.76.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Williams J. G., Lloyd M. M. Changes in the abundance of polyadenylated RNA during slime mould development measured using cloned molecular hybridization probes. J Mol Biol. 1979 Mar 25;129(1):19–35. doi: 10.1016/0022-2836(79)90056-1. [DOI] [PubMed] [Google Scholar]
  36. de Wet J. R., Daniels D. L., Schroeder J. L., Williams B. G., Denniston-Thompson K., Moore D. D., Blattner F. R. Restriction maps for twenty-one Charon vector phages. J Virol. 1980 Jan;33(1):401–410. doi: 10.1128/jvi.33.1.401-410.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES