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Nucleic Acids Res. 1994 Aug 25; 22(16): 3339–3346.
doi: 10.1093/nar/22.16.3339
PMCID: PMC523727
PMID: 8078769

DNA binding and bending properties of the post-meiotically expressed Sry-related protein Sox-5.

F Connor, P D Cary, C M Read, N S Preston, P C Driscoll, P Denny, C Crane-Robinson, and A Ashworth
Author information Copyright and License information Disclaimer

Abstract

Sox-5 is one of a family of genes which show homology to the HMG box region of the testis determining gene SRY. We have used indirect immunofluorescence to show that Sox-5 protein is localized to the nucleus of post-meiotic round spermatids in the mouse testis. In vitro footprinting and gel retardation assays demonstrate that Sox-5 binds specifically to the sequence AACAAT with moderately high affinity (Kd of approximately 10(-9) M). Moreover, interaction of Sox-5 with its target DNA induces a significant bend in the DNA, characteristic of HMG box proteins. Circular dichroism spectroscopy of the Sox-5 HMG box and its specific complex with DNA shows an alteration in the DNA spectrum, perhaps as a consequence of DNA bending, but none in the protein spectrum on complex formation. The dependence of the change in the CD spectrum with protein to DNA ratio demonstrates the formation of a 1:1 complex. Analysis of the structure of the Sox-5 HMG box by 2D NMR suggests that both the location of helical secondary structure as well as the tertiary structure is similar to that of HMG1 box 2.

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Selected References

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  • 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. [PubMed] [Google Scholar]
  • Laudet V, Stehelin D, Clevers H. Ancestry and diversity of the HMG box superfamily. Nucleic Acids Res. 1993 May 25;21(10):2493–2501. [PMC free article] [PubMed] [Google Scholar]
  • 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. [PMC free article] [PubMed] [Google Scholar]
  • 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. [PubMed] [Google Scholar]
  • 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. [PMC free article] [PubMed] [Google Scholar]
  • Staben C, Yanofsky C. Neurospora crassa a mating-type region. Proc Natl Acad Sci U S A. 1990 Jul;87(13):4917–4921. [PMC free article] [PubMed] [Google Scholar]
  • 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. [PubMed] [Google Scholar]
  • Denny P, Swift S, Brand N, Dabhade N, Barton P, Ashworth A. A conserved family of genes related to the testis determining gene, SRY. Nucleic Acids Res. 1992 Jun 11;20(11):2887–2887. [PMC free article] [PubMed] [Google Scholar]
  • 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. [PMC free article] [PubMed] [Google Scholar]
  • Willison K, Lewis V, Zuckerman KS, Cordell J, Dean C, Miller K, Lyon MF, Marsh M. The t complex polypeptide 1 (TCP-1) is associated with the cytoplasmic aspect of Golgi membranes. Cell. 1989 May 19;57(4):621–632. [PubMed] [Google Scholar]
  • Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. [PubMed] [Google Scholar]
  • Studier FW. Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. J Mol Biol. 1991 May 5;219(1):37–44. [PubMed] [Google Scholar]
  • Read CM, Cary PD, Crane-Robinson C, Driscoll PC, Norman DG. Solution structure of a DNA-binding domain from HMG1. Nucleic Acids Res. 1993 Jul 25;21(15):3427–3436. [PMC free article] [PubMed] [Google Scholar]
  • Liu-Johnson HN, Gartenberg MR, Crothers DM. The DNA binding domain and bending angle of E. coli CAP protein. Cell. 1986 Dec 26;47(6):995–1005. [PubMed] [Google Scholar]
  • Kim J, Zwieb C, Wu C, Adhya S. Bending of DNA by gene-regulatory proteins: construction and use of a DNA bending vector. Gene. 1989 Dec 21;85(1):15–23. [PubMed] [Google Scholar]
  • Thompson JF, Landy A. Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. Nucleic Acids Res. 1988 Oct 25;16(20):9687–9705. [PMC free article] [PubMed] [Google Scholar]
  • Nasrin N, Buggs C, Kong XF, 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. [PubMed] [Google Scholar]
  • Harley VR, Jackson DI, Hextall PJ, Hawkins JR, Berkovitz GD, Sockanathan S, Lovell-Badge R, Goodfellow PN. DNA binding activity of recombinant SRY from normal males and XY females. Science. 1992 Jan 24;255(5043):453–456. [PubMed] [Google Scholar]
  • 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. [PubMed] [Google Scholar]
  • Harley VR, Lovell-Badge R, Goodfellow PN. Definition of a consensus DNA binding site for SRY. Nucleic Acids Res. 1994 Apr 25;22(8):1500–1501. [PMC free article] [PubMed] [Google Scholar]
  • Giese K, Pagel J, Grosschedl R. Distinct DNA-binding properties of the high mobility group domain of murine and human SRY sex-determining factors. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3368–3372. [PMC free article] [PubMed] [Google Scholar]
  • Giese K, Amsterdam A, Grosschedl R. DNA-binding properties of the HMG domain of the lymphoid-specific transcriptional regulator LEF-1. Genes Dev. 1991 Dec;5(12B):2567–2578. [PubMed] [Google Scholar]
  • 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. [PMC free article] [PubMed] [Google Scholar]
  • Lilley DM. DNA--protein interactions. HMG has DNA wrapped up. Nature. 1992 May 28;357(6376):282–283. [PubMed] [Google Scholar]
  • Wu HM, Crothers DM. The locus of sequence-directed and protein-induced DNA bending. Nature. 1984 Apr 5;308(5959):509–513. [PubMed] [Google Scholar]
  • Kerppola TK, Curran T. DNA bending by Fos and Jun: the flexible hinge model. Science. 1991 Nov 22;254(5035):1210–1214. [PubMed] [Google Scholar]
  • Ferrari S, Harley VR, Pontiggia A, Goodfellow PN, Lovell-Badge R, Bianchi ME. SRY, like HMG1, recognizes sharp angles in DNA. EMBO J. 1992 Dec;11(12):4497–4506. [PMC free article] [PubMed] [Google Scholar]
  • Olins DE, Bryan PN, Harrington RE, Hill WE, Olins AL. Conformational states of chromatin nu bodies induced by urea. Nucleic Acids Res. 1977 Jun;4(6):1911–1931. [PMC free article] [PubMed] [Google Scholar]
  • Compton LA, Johnson WC., Jr Analysis of protein circular dichroism spectra for secondary structure using a simple matrix multiplication. Anal Biochem. 1986 May 15;155(1):155–167. [PubMed] [Google Scholar]
  • van Houte L, van Oers A, van de Wetering M, Dooijes D, Kaptein R, Clevers H. The sequence-specific high mobility group 1 box of TCF-1 adopts a predominantly alpha-helical conformation in solution. J Biol Chem. 1993 Aug 25;268(24):18083–18087. [PubMed] [Google Scholar]
  • Weir HM, Kraulis PJ, Hill CS, Raine AR, Laue ED, Thomas JO. Structure of the HMG box motif in the B-domain of HMG1. EMBO J. 1993 Apr;12(4):1311–1319. [PMC free article] [PubMed] [Google Scholar]
  • Ner SS. HMGs everywhere. Curr Biol. 1992 Apr;2(4):208–210. [PubMed] [Google Scholar]
  • McLaren A. Sex determination in mammals. Trends Genet. 1988 Jun;4(6):153–157. [PubMed] [Google Scholar]
  • 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. [PubMed] [Google Scholar]
  • Jantzen HM, Admon A, Bell SP, Tjian R. Nucleolar transcription factor hUBF contains a DNA-binding motif with homology to HMG proteins. Nature. 1990 Apr 26;344(6269):830–836. [PubMed] [Google Scholar]
  • Sinclair AH, Berta P, Palmer MS, Hawkins JR, Griffiths BL, Smith MJ, Foster JW, Frischauf AM, Lovell-Badge R, Goodfellow PN. 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. [PubMed] [Google Scholar]
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