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. 1995 Mar 1;128(5):737–748. doi: 10.1083/jcb.128.5.737

Nuclear localization of the testis determining gene product SRY

PMCID: PMC2120386  PMID: 7876301

Abstract

We have studied the expression of the human SRY protein (termed p27SRY) in two different cell lines by using specific antibodies. Confocal microscopy enabled us to localize p27SRY precisely in the nucleus in a discrete punctuate pattern. Furthermore, through microinjection experiments, we have demonstrated that the localization of the p27SRY protein into the nucleus was an event involving the NH2-terminal part of the high mobility group (HMG) domain. With the help of several synthetic peptides and various p27SRY mutants, we have characterized a bipartite basic motif in this part of the protein corresponding to a nuclear localization signal. This nuclear localization signal appears to be highly conserved in SRY box- and HMB box-containing proteins, suggesting common properties of nuclear targeting within the HMG box protein family.

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Selected References

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  1. Andrews P. W., Damjanov I., Simon D., Banting G. S., Carlin C., Dracopoli N. C., Føgh J. Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. Lab Invest. 1984 Feb;50(2):147–162. [PubMed] [Google Scholar]
  2. Behlke M. A., Bogan J. S., Beer-Romero P., Page D. C. Evidence that the SRY protein is encoded by a single exon on the human Y chromosome. Genomics. 1993 Sep;17(3):736–739. doi: 10.1006/geno.1993.1395. [DOI] [PubMed] [Google Scholar]
  3. Bell S. P., Jantzen H. M., Tjian R. Assembly of alternative multiprotein complexes directs rRNA promoter selectivity. Genes Dev. 1990 Jun;4(6):943–954. doi: 10.1101/gad.4.6.943. [DOI] [PubMed] [Google Scholar]
  4. Berta P., Hawkins J. R., Sinclair A. H., Taylor A., Griffiths B. L., Goodfellow P. N., Fellous M. Genetic evidence equating SRY and the testis-determining factor. Nature. 1990 Nov 29;348(6300):448–450. doi: 10.1038/348448A0. [DOI] [PubMed] [Google Scholar]
  5. Clépet C., Schafer A. J., Sinclair A. H., Palmer M. S., Lovell-Badge R., Goodfellow P. N. The human SRY transcript. Hum Mol Genet. 1993 Dec;2(12):2007–2012. doi: 10.1093/hmg/2.12.2007. [DOI] [PubMed] [Google Scholar]
  6. Denny P., Swift S., Connor F., Ashworth A. An SRY-related gene expressed during spermatogenesis in the mouse encodes a sequence-specific DNA-binding protein. EMBO J. 1992 Oct;11(10):3705–3712. doi: 10.1002/j.1460-2075.1992.tb05455.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dingwall C., Laskey R. A. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. doi: 10.1016/0968-0004(91)90184-w. [DOI] [PubMed] [Google Scholar]
  8. Dingwall C., Robbins J., Dilworth S. M., Roberts B., Richardson W. D. The nucleoplasmin nuclear location sequence is larger and more complex than that of SV-40 large T antigen. J Cell Biol. 1988 Sep;107(3):841–849. doi: 10.1083/jcb.107.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fernandez A., Mery J., Vandromme M., Basset M., Cavadore J. C., Lamb N. J. Effective intracellular inhibition of the cAMP-dependent protein kinase by microinjection of a modified form of the specific inhibitor peptide PKi in living fibroblasts. Exp Cell Res. 1991 Aug;195(2):468–477. doi: 10.1016/0014-4827(91)90398-e. [DOI] [PubMed] [Google Scholar]
  10. Ferrari S., Harley V. R., Pontiggia A., Goodfellow P. N., Lovell-Badge R., Bianchi M. E. SRY, like HMG1, recognizes sharp angles in DNA. EMBO J. 1992 Dec;11(12):4497–4506. doi: 10.1002/j.1460-2075.1992.tb05551.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Foster J. W., Brennan F. E., Hampikian G. K., Goodfellow P. N., Sinclair A. H., Lovell-Badge R., Selwood L., Renfree M. B., Cooper D. W., Graves J. A. Evolution of sex determination and the Y chromosome: SRY-related sequences in marsupials. Nature. 1992 Oct 8;359(6395):531–533. doi: 10.1038/359531a0. [DOI] [PubMed] [Google Scholar]
  12. Giese K., Cox J., Grosschedl R. The HMG domain of lymphoid enhancer factor 1 bends DNA and facilitates assembly of functional nucleoprotein structures. Cell. 1992 Apr 3;69(1):185–195. doi: 10.1016/0092-8674(92)90129-z. [DOI] [PubMed] [Google Scholar]
  13. Giese K., Grosschedl R. LEF-1 contains an activation domain that stimulates transcription only in a specific context of factor-binding sites. EMBO J. 1993 Dec;12(12):4667–4676. doi: 10.1002/j.1460-2075.1993.tb06155.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Griess E. A., Rensing S. A., Grasser K. D., Maier U. G., Feix G. Phylogenetic relationships of HMG box DNA-binding domains. J Mol Evol. 1993 Aug;37(2):204–210. doi: 10.1007/BF02407357. [DOI] [PubMed] [Google Scholar]
  15. Griffiths R. The isolation of conserved DNA sequences related to the human sex-determining region Y gene from the lesser black-backed gull (Larus fuscus). Proc Biol Sci. 1991 May 22;244(1310):123–128. doi: 10.1098/rspb.1991.0060. [DOI] [PubMed] [Google Scholar]
  16. Gubbay J., Collignon J., Koopman P., Capel B., Economou A., Münsterberg A., Vivian N., Goodfellow P., Lovell-Badge R. A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature. 1990 Jul 19;346(6281):245–250. doi: 10.1038/346245a0. [DOI] [PubMed] [Google Scholar]
  17. Harley V. R., Jackson D. I., Hextall P. J., Hawkins J. R., Berkovitz G. D., Sockanathan S., Lovell-Badge R., Goodfellow P. N. DNA binding activity of recombinant SRY from normal males and XY females. Science. 1992 Jan 24;255(5043):453–456. doi: 10.1126/science.1734522. [DOI] [PubMed] [Google Scholar]
  18. Hawkins J. R., Taylor A., Berta P., Levilliers J., Van der Auwera B., Goodfellow P. N. Mutational analysis of SRY: nonsense and missense mutations in XY sex reversal. Hum Genet. 1992 Feb;88(4):471–474. doi: 10.1007/BF00215684. [DOI] [PubMed] [Google Scholar]
  19. Härd T., Kellenbach E., Boelens R., Maler B. A., Dahlman K., Freedman L. P., Carlstedt-Duke J., Yamamoto K. R., Gustafsson J. A., Kaptein R. Solution structure of the glucocorticoid receptor DNA-binding domain. Science. 1990 Jul 13;249(4965):157–160. doi: 10.1126/science.2115209. [DOI] [PubMed] [Google Scholar]
  20. Josso N., Cate R. L., Picard J. Y., Vigier B., di Clemente N., Wilson C., Imbeaud S., Pepinsky R. B., Guerrier D., Boussin L. Anti-müllerian hormone: the Jost factor. Recent Prog Horm Res. 1993;48:1–59. doi: 10.1016/b978-0-12-571148-7.50005-1. [DOI] [PubMed] [Google Scholar]
  21. Jost A., Magre S., Agelopoulou R. Early stages of testicular differentiation in the rat. Hum Genet. 1981;58(1):59–63. doi: 10.1007/BF00284150. [DOI] [PubMed] [Google Scholar]
  22. Jäger R. J., Anvret M., Hall K., Scherer G. A human XY female with a frame shift mutation in the candidate testis-determining gene SRY. Nature. 1990 Nov 29;348(6300):452–454. doi: 10.1038/348452a0. [DOI] [PubMed] [Google Scholar]
  23. Kelly M., Burke J., Smith M., Klar A., Beach D. Four mating-type genes control sexual differentiation in the fission yeast. EMBO J. 1988 May;7(5):1537–1547. doi: 10.1002/j.1460-2075.1988.tb02973.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koopman P., Gubbay J., Vivian N., Goodfellow P., Lovell-Badge R. Male development of chromosomally female mice transgenic for Sry. Nature. 1991 May 9;351(6322):117–121. doi: 10.1038/351117a0. [DOI] [PubMed] [Google Scholar]
  25. Koopman P., Münsterberg A., Capel B., Vivian N., Lovell-Badge R. Expression of a candidate sex-determining gene during mouse testis differentiation. Nature. 1990 Nov 29;348(6300):450–452. doi: 10.1038/348450a0. [DOI] [PubMed] [Google Scholar]
  26. McElreavey K., Vilain E., Abbas N., Herskowitz I., Fellous M. A regulatory cascade hypothesis for mammalian sex determination: SRY represses a negative regulator of male development. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3368–3372. doi: 10.1073/pnas.90.8.3368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nasrin N., Buggs C., Kong X. F., Carnazza J., Goebl M., Alexander-Bridges M. DNA-binding properties of the product of the testis-determining gene and a related protein. Nature. 1991 Nov 28;354(6351):317–320. doi: 10.1038/354317a0. [DOI] [PubMed] [Google Scholar]
  28. Ner S. S. HMGs everywhere. Curr Biol. 1992 Apr;2(4):208–210. doi: 10.1016/0960-9822(92)90541-h. [DOI] [PubMed] [Google Scholar]
  29. Nicola N. A., Metcalf D. Subunit promiscuity among hemopoietic growth factor receptors. Cell. 1991 Oct 4;67(1):1–4. doi: 10.1016/0092-8674(91)90564-f. [DOI] [PubMed] [Google Scholar]
  30. Peters R. Fluorescence microphotolysis to measure nucleocytoplasmic transport and intracellular mobility. Biochim Biophys Acta. 1986 Dec 22;864(3-4):305–359. doi: 10.1016/0304-4157(86)90003-1. [DOI] [PubMed] [Google Scholar]
  31. Peters R. Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis. EMBO J. 1984 Aug;3(8):1831–1836. doi: 10.1002/j.1460-2075.1984.tb02055.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Poulat F., Guichard G., Gozé C., Heitz F., Calas B., Berta P. Synthesis of a large peptide mimicking the DNA binding properties of the sex determining protein, SRY. FEBS Lett. 1992 Sep 14;309(3):385–388. doi: 10.1016/0014-5793(92)80812-u. [DOI] [PubMed] [Google Scholar]
  33. Poulat F., Soullier S., Gozé C., Heitz F., Calas B., Berta P. Description and functional implications of a novel mutation in the sex-determining gene SRY. Hum Mutat. 1994;3(3):200–204. doi: 10.1002/humu.1380030305. [DOI] [PubMed] [Google Scholar]
  34. Shatzman A. R., Rosenberg M. Expression, identification, and characterization of recombinant gene products in Escherichia coli. Methods Enzymol. 1987;152:661–673. doi: 10.1016/0076-6879(87)52072-9. [DOI] [PubMed] [Google Scholar]
  35. Sinclair A. H., Berta P., Palmer M. S., Hawkins J. R., Griffiths B. L., Smith M. J., Foster J. W., Frischauf A. M., Lovell-Badge R., Goodfellow P. N. A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif. Nature. 1990 Jul 19;346(6281):240–244. doi: 10.1038/346240a0. [DOI] [PubMed] [Google Scholar]
  36. Su H., Lau Y. F. Identification of the transcriptional unit, structural organization, and promoter sequence of the human sex-determining region Y (SRY) gene, using a reverse genetic approach. Am J Hum Genet. 1993 Jan;52(1):24–38. [PMC free article] [PubMed] [Google Scholar]
  37. TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Travis A., Amsterdam A., Belanger C., Grosschedl R. LEF-1, a gene encoding a lymphoid-specific protein with an HMG domain, regulates T-cell receptor alpha enhancer function [corrected]. Genes Dev. 1991 May;5(5):880–894. doi: 10.1101/gad.5.5.880. [DOI] [PubMed] [Google Scholar]
  39. Tricoli J. V., Yao J. L., D'Souza S. A., Bracken R. B. Detection of sex-region Y (SRY) transcripts in human prostate adenocarcinoma and benign prostatic hypertrophy. Genes Chromosomes Cancer. 1993 Sep;8(1):28–33. doi: 10.1002/gcc.2870080106. [DOI] [PubMed] [Google Scholar]
  40. Tucker P. K., Lundrigan B. L. Rapid evolution of the sex determining locus in Old World mice and rats. Nature. 1993 Aug 19;364(6439):715–717. doi: 10.1038/364715a0. [DOI] [PubMed] [Google Scholar]
  41. Wen L., Huang J. K., Johnson B. H., Reeck G. R. A human placental cDNA clone that encodes nonhistone chromosomal protein HMG-1. Nucleic Acids Res. 1989 Feb 11;17(3):1197–1214. doi: 10.1093/nar/17.3.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Whitfield L. S., Lovell-Badge R., Goodfellow P. N. Rapid sequence evolution of the mammalian sex-determining gene SRY. Nature. 1993 Aug 19;364(6439):713–715. doi: 10.1038/364713a0. [DOI] [PubMed] [Google Scholar]
  43. van de Wetering M., Clevers H. Sequence-specific interaction of the HMG box proteins TCF-1 and SRY occurs within the minor groove of a Watson-Crick double helix. EMBO J. 1992 Aug;11(8):3039–3044. doi: 10.1002/j.1460-2075.1992.tb05374.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. van de Wetering M., Oosterwegel M., Dooijes D., Clevers H. Identification and cloning of TCF-1, a T lymphocyte-specific transcription factor containing a sequence-specific HMG box. EMBO J. 1991 Jan;10(1):123–132. doi: 10.1002/j.1460-2075.1991.tb07928.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. van de Wetering M., Oosterwegel M., van Norren K., Clevers H. Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes. EMBO J. 1993 Oct;12(10):3847–3854. doi: 10.1002/j.1460-2075.1993.tb06063.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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