Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1998 Apr 1;26(7):1675–1680. doi: 10.1093/nar/26.7.1675

Dimerization of the testis brain RNA-binding protein (translin) is mediated through its C-terminus and is required for DNA- and RNA-binding.

X Q Wu 1, L Xu 1, N B Hecht 1
PMCID: PMC147482  PMID: 9512538

Abstract

Testis brain-RNA-binding protein (TB-RBP) is a single-stranded DNA- and RNA-binding protein that is involved in chromosomal translocations, mRNA transport and translational regulation. Here we show from in vitro and in vivo protein binding studies that TB-RBP dimers are the minimum structural unit needed for DNA- and RNA-binding. Truncation studies demonstrate that the C-terminus of 55 amino acids of TB-RBP is essential, but not sufficient for DNA- or RNA-binding, and deletion of the leucine zipper motif in the C-terminus abolishes DNA- and RNA-binding. Changing cysteine 225 in the C-terminus to alanine does not significantly reduce DNA- or RNA-binding, but reduces the stability of the dimer. We conclude that the leucine zipper motif is required to maintain two molecules of TB-RBP as a dimer which is stabilized by a disulfide bond involving cysteine 225.

Full Text

The Full Text of this article is available as a PDF (291.9 KB).

Selected References

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

  1. Amati B., Littlewood T. D., Evan G. I., Land H. The c-Myc protein induces cell cycle progression and apoptosis through dimerization with Max. EMBO J. 1993 Dec 15;12(13):5083–5087. doi: 10.1002/j.1460-2075.1993.tb06202.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aoki K., Inazawa J., Takahashi T., Nakahara K., Kasai M. Genomic structure and chromosomal localization of the gene encoding translin, a recombination hotspot binding protein. Genomics. 1997 Jul 15;43(2):237–241. doi: 10.1006/geno.1997.4796. [DOI] [PubMed] [Google Scholar]
  3. Aoki K., Suzuki K., Sugano T., Tasaka T., Nakahara K., Kuge O., Omori A., Kasai M. A novel gene, Translin, encodes a recombination hotspot binding protein associated with chromosomal translocations. Nat Genet. 1995 Jun;10(2):167–174. doi: 10.1038/ng0695-167. [DOI] [PubMed] [Google Scholar]
  4. Benbrook D. M., Jones N. C. Heterodimer formation between CREB and JUN proteins. Oncogene. 1990 Mar;5(3):295–302. [PubMed] [Google Scholar]
  5. Busch S. J., Sassone-Corsi P. Dimers, leucine zippers and DNA-binding domains. Trends Genet. 1990 Feb;6(2):36–40. doi: 10.1016/0168-9525(90)90071-d. [DOI] [PubMed] [Google Scholar]
  6. Grandori C., Eisenman R. N. Myc target genes. Trends Biochem Sci. 1997 May;22(5):177–181. doi: 10.1016/s0968-0004(97)01025-6. [DOI] [PubMed] [Google Scholar]
  7. Gutman A., Wasylyk B. Nuclear targets for transcription regulation by oncogenes. Trends Genet. 1991 Feb;7(2):49–54. doi: 10.1016/0168-9525(91)90231-E. [DOI] [PubMed] [Google Scholar]
  8. Hai T., Curran T. Cross-family dimerization of transcription factors Fos/Jun and ATF/CREB alters DNA binding specificity. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3720–3724. doi: 10.1073/pnas.88.9.3720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Han J. R., Gu W., Hecht N. B. Testis-brain RNA-binding protein, a testicular translational regulatory RNA-binding protein, is present in the brain and binds to the 3' untranslated regions of transported brain mRNAs. Biol Reprod. 1995 Sep;53(3):707–717. doi: 10.1095/biolreprod53.3.707. [DOI] [PubMed] [Google Scholar]
  10. Han J. R., Yiu G. K., Hecht N. B. Testis/brain RNA-binding protein attaches translationally repressed and transported mRNAs to microtubules. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9550–9554. doi: 10.1073/pnas.92.21.9550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hope I. A., Struhl K. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA. EMBO J. 1987 Sep;6(9):2781–2784. doi: 10.1002/j.1460-2075.1987.tb02573.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kasai M., Matsuzaki T., Katayanagi K., Omori A., Maziarz R. T., Strominger J. L., Aoki K., Suzuki K. The translin ring specifically recognizes DNA ends at recombination hot spots in the human genome. J Biol Chem. 1997 Apr 25;272(17):11402–11407. doi: 10.1074/jbc.272.17.11402. [DOI] [PubMed] [Google Scholar]
  13. Kwon Y. K., Hecht N. B. Binding of a phosphoprotein to the 3' untranslated region of the mouse protamine 2 mRNA temporally represses its translation. Mol Cell Biol. 1993 Oct;13(10):6547–6557. doi: 10.1128/mcb.13.10.6547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kwon Y. K., Hecht N. B. Cytoplasmic protein binding to highly conserved sequences in the 3' untranslated region of mouse protamine 2 mRNA, a translationally regulated transcript of male germ cells. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3584–3588. doi: 10.1073/pnas.88.9.3584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Landschulz W. H., Johnson P. F., McKnight S. L. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988 Jun 24;240(4860):1759–1764. doi: 10.1126/science.3289117. [DOI] [PubMed] [Google Scholar]
  17. MacDougald O. A., Lane M. D. Transcriptional regulation of gene expression during adipocyte differentiation. Annu Rev Biochem. 1995;64:345–373. doi: 10.1146/annurev.bi.64.070195.002021. [DOI] [PubMed] [Google Scholar]
  18. McAfee J. G., Shahied-Milam L., Soltaninassab S. R., LeStourgeon W. M. A major determinant of hnRNP C protein binding to RNA is a novel bZIP-like RNA binding domain. RNA. 1996 Nov;2(11):1139–1152. [PMC free article] [PubMed] [Google Scholar]
  19. Suck D. Common fold, common function, common origin? Nat Struct Biol. 1997 Mar;4(3):161–165. doi: 10.1038/nsb0397-161. [DOI] [PubMed] [Google Scholar]
  20. Wu X. Q., Gu W., Meng X., Hecht N. B. The RNA-binding protein, TB-RBP, is the mouse homologue of translin, a recombination protein associated with chromosomal translocations. Proc Natl Acad Sci U S A. 1997 May 27;94(11):5640–5645. doi: 10.1073/pnas.94.11.5640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yurkova M. S., Murray M. T. A translation regulatory particle containing the Xenopus oocyte Y box protein mRNP3+4. J Biol Chem. 1997 Apr 18;272(16):10870–10876. doi: 10.1074/jbc.272.16.10870. [DOI] [PubMed] [Google Scholar]

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

RESOURCES