Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1997 Apr;71(4):2873–2880. doi: 10.1128/jvi.71.4.2873-2880.1997

Chicken YB-2, a Y-box protein, is a potent activator of Rous sarcoma virus long terminal repeat-driven transcription in avian fibroblasts.

S K Swamynathan 1, A Nambiar 1, R V Guntaka 1
PMCID: PMC191413  PMID: 9060644

Abstract

We have previously reported on the cloning and characterization of chk-YB-2, a novel member of the Y-box family of proteins, that binds to the sequence 5'-GTACCACC-3' present on the noncoding strand of the Rous sarcoma virus (RSV) long terminal repeat (LTR) in a single-strand-specific manner. Here, we demonstrate that deletion or mutation of this motif not only eliminates chk-YB-2 binding in vitro but also down-regulates RSV LTR-driven transcription in avian cells. Selective abrogation of chk-YB-2 expression by using antisense oligonucleotides decreased RSV LTR-driven transcription in a promoter-specific manner. This inhibition was not observed when a reporter construct with a deletion in the chk-YB-2 binding site was used. Depletion of cellular chk-YB-2 by transfecting the cells with excess of its recognition sequence oligonucleotides also resulted in reduced transcription from the RSV LTR. Taken together, these results suggest that chk-YB-2 acts as an activator of LTR-promoted transcription in avian cells and that this activation is mediated primarily through the sequence 5'-GTACCACC-3'.

Full Text

The Full Text of this article is available as a PDF (805.7 KB).

Selected References

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

  1. Abdulkadir S. A., Ono S. J. How are class II MHC genes turned on and off? FASEB J. 1995 Nov;9(14):1429–1435. doi: 10.1096/fasebj.9.14.7589984. [DOI] [PubMed] [Google Scholar]
  2. Boulden A., Sealy L. Identification of a third protein factor which binds to the Rous sarcoma virus LTR enhancer: possible homology with the serum response factor. Virology. 1990 Jan;174(1):204–216. doi: 10.1016/0042-6822(90)90069-4. [DOI] [PubMed] [Google Scholar]
  3. Cleavinger P. J., Shin B. A., Kandala J. C., Nambiar A., Swamynathan S. K., Guntaka R. V. Cloning of Rous sarcoma virus enhancer factor genes. II. RSV-EF-II, abundantly expressed in fibroblasts and muscle tissue, binds to an octamer sequence, 5'-GTACCACC-3', in the noncoding strand of RSV enhancer. Virology. 1996 Aug 1;222(1):133–143. doi: 10.1006/viro.1996.0404. [DOI] [PubMed] [Google Scholar]
  4. Cohen I., Reynolds W. F. The Xenopus YB3 protein binds the B box element of the class III promoter. Nucleic Acids Res. 1991 Sep 11;19(17):4753–4759. doi: 10.1093/nar/19.17.4753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cullen B. R., Raymond K., Ju G. Functional analysis of the transcription control region located within the avian retroviral long terminal repeat. Mol Cell Biol. 1985 Mar;5(3):438–447. doi: 10.1128/mcb.5.3.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cullen B. R., Raymond K., Ju G. Transcriptional activity of avian retroviral long terminal repeats directly correlates with enhancer activity. J Virol. 1985 Feb;53(2):515–521. doi: 10.1128/jvi.53.2.515-521.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Didier D. K., Schiffenbauer J., Woulfe S. L., Zacheis M., Schwartz B. D. Characterization of the cDNA encoding a protein binding to the major histocompatibility complex class II Y box. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7322–7326. doi: 10.1073/pnas.85.19.7322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dutta A., Stoeckle M. Y., Hanafusa H. Serum and v-src increase the level of a CCAAT-binding factor required for transcription from a retroviral long terminal repeat. Genes Dev. 1990 Feb;4(2):243–254. doi: 10.1101/gad.4.2.243. [DOI] [PubMed] [Google Scholar]
  9. Gai X. X., Lipson K. E., Prystowsky M. B. Unusual DNA binding characteristics of an in vitro translation product of the CCAAT binding protein mYB-1. Nucleic Acids Res. 1992 Feb 11;20(3):601–606. doi: 10.1093/nar/20.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goodwin G. H. Identification of three sequence-specific DNA-binding proteins which interact with the Rous sarcoma virus enhancer and upstream promoter elements. J Virol. 1988 Jun;62(6):2186–2190. doi: 10.1128/jvi.62.6.2186-2190.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gowda S., Rao A. S., Kim Y. W., Guntaka R. V. Identification of sequences in the long terminal repeat of avian sarcoma virus required for efficient transcription. Virology. 1988 Jan;162(1):243–247. doi: 10.1016/0042-6822(88)90415-1. [DOI] [PubMed] [Google Scholar]
  14. Grant C. E., Deeley R. G. Cloning and characterization of chicken YB-1: regulation of expression in the liver. Mol Cell Biol. 1993 Jul;13(7):4186–4196. doi: 10.1128/mcb.13.7.4186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Greuel B. T., Sealy L., Majors J. E. Transcriptional activity of the Rous sarcoma virus long terminal repeat correlates with binding of a factor to an upstream CCAAT box in vitro. Virology. 1990 Jul;177(1):33–43. doi: 10.1016/0042-6822(90)90457-3. [DOI] [PubMed] [Google Scholar]
  16. Houtz E. K., Conklin K. F. Identification of EFIV, a stable factor present in many avian cell types that transactivates sequences in the 5' portion of the Rous sarcoma virus long terminal repeat enhancer. J Virol. 1996 Jan;70(1):393–401. doi: 10.1128/jvi.70.1.393-401.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kandala J. C., Guntaka R. V. Cloning of Rous sarcoma virus enhancer factor genes. I. Evidence that RSV-EF-I is related to Y-box (inverted CCAAT) binding proteins and binds to multiple motifs in the RSV enhancer. Virology. 1994 Feb;198(2):514–523. doi: 10.1006/viro.1994.1062. [DOI] [PubMed] [Google Scholar]
  18. Kenny S., Guntaka R. V. Localization by mutational analysis of transcription factor binding sequences in the U3 region of Rous sarcoma virus LTR. Virology. 1990 Jun;176(2):483–493. doi: 10.1016/0042-6822(90)90018-m. [DOI] [PubMed] [Google Scholar]
  19. Kolluri R., Torrey T. A., Kinniburgh A. J. A CT promoter element binding protein: definition of a double-strand and a novel single-strand DNA binding motif. Nucleic Acids Res. 1992 Jan 11;20(1):111–116. doi: 10.1093/nar/20.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ladomery M., Sommerville J. A role for Y-box proteins in cell proliferation. Bioessays. 1995 Jan;17(1):9–11. doi: 10.1002/bies.950170104. [DOI] [PubMed] [Google Scholar]
  21. Laimins L. A., Tsichlis P., Khoury G. Multiple enhancer domains in the 3' terminus of the Prague strain of Rous sarcoma virus. Nucleic Acids Res. 1984 Aug 24;12(16):6427–6442. doi: 10.1093/nar/12.16.6427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MacDonald G. H., Itoh-Lindstrom Y., Ting J. P. The transcriptional regulatory protein, YB-1, promotes single-stranded regions in the DRA promoter. J Biol Chem. 1995 Feb 24;270(8):3527–3533. doi: 10.1074/jbc.270.8.3527. [DOI] [PubMed] [Google Scholar]
  23. Majors J. The structure and function of retroviral long terminal repeats. Curr Top Microbiol Immunol. 1990;157:49–92. doi: 10.1007/978-3-642-75218-6_3. [DOI] [PubMed] [Google Scholar]
  24. Mitsialis S. A., Manley J. L., Guntaka R. V. Localization of active promoters for eucaryotic RNA polymerase II in the long terminal repeat of avian sarcoma virus DNA. Mol Cell Biol. 1983 May;3(5):811–818. doi: 10.1128/mcb.3.5.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Norton P. A., Coffin J. M. Characterization of Rous sarcoma virus sequences essential for viral gene expression. J Virol. 1987 Apr;61(4):1171–1179. doi: 10.1128/jvi.61.4.1171-1179.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ozer J., Faber M., Chalkley R., Sealy L. Isolation and characterization of a cDNA clone for the CCAAT transcription factor EFIA reveals a novel structural motif. J Biol Chem. 1990 Dec 25;265(36):22143–22152. [PubMed] [Google Scholar]
  27. Raj G. V., Safak M., MacDonald G. H., Khalili K. Transcriptional regulation of human polyomavirus JC: evidence for a functional interaction between RelA (p65) and the Y-box-binding protein, YB-1. J Virol. 1996 Sep;70(9):5944–5953. doi: 10.1128/jvi.70.9.5944-5953.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ranjan M., Tafuri S. R., Wolffe A. P. Masking mRNA from translation in somatic cells. Genes Dev. 1993 Sep;7(9):1725–1736. doi: 10.1101/gad.7.9.1725. [DOI] [PubMed] [Google Scholar]
  29. Ruddell A., Linial M. L., Groudine M. Tissue-specific lability and expression of avian leukosis virus long terminal repeat enhancer-binding proteins. Mol Cell Biol. 1989 Dec;9(12):5660–5668. doi: 10.1128/mcb.9.12.5660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ruddell A. Transcription regulatory elements of the avian retroviral long terminal repeat. Virology. 1995 Jan 10;206(1):1–7. doi: 10.1016/s0042-6822(95)80013-1. [DOI] [PubMed] [Google Scholar]
  31. Ryden T. A., de Mars M., Beemon K. Mutation of the C/EBP binding sites in the Rous sarcoma virus long terminal repeat and gag enhancers. J Virol. 1993 May;67(5):2862–2870. doi: 10.1128/jvi.67.5.2862-2870.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sealey L., Chalkley R. At least two nuclear proteins bind specifically to the Rous sarcoma virus long terminal repeat enhancer. Mol Cell Biol. 1987 Feb;7(2):787–798. doi: 10.1128/mcb.7.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sears R. C., Sealy L. Characterization of nuclear proteins that bind the EFII enhancer sequence in the Rous sarcoma virus long terminal repeat. J Virol. 1992 Nov;66(11):6338–6352. doi: 10.1128/jvi.66.11.6338-6352.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sommerville J., Ladomery M. Masking of mRNA by Y-box proteins. FASEB J. 1996 Mar;10(4):435–443. doi: 10.1096/fasebj.10.4.8647342. [DOI] [PubMed] [Google Scholar]
  35. Tafuri S. R., Familari M., Wolffe A. P. A mouse Y box protein, MSY1, is associated with paternal mRNA in spermatocytes. J Biol Chem. 1993 Jun 5;268(16):12213–12220. [PubMed] [Google Scholar]
  36. Tafuri S. R., Wolffe A. P. DNA binding, multimerization, and transcription stimulation by the Xenopus Y box proteins in vitro. New Biol. 1992 Apr;4(4):349–359. [PubMed] [Google Scholar]
  37. Tafuri S. R., Wolffe A. P. Selective recruitment of masked maternal mRNA from messenger ribonucleoprotein particles containing FRGY2 (mRNP4). J Biol Chem. 1993 Nov 15;268(32):24255–24261. [PubMed] [Google Scholar]
  38. Ting J. P., Painter A., Zeleznik-Le N. J., MacDonald G., Moore T. M., Brown A., Schwartz B. D. YB-1 DNA-binding protein represses interferon gamma activation of class II major histocompatibility complex genes. J Exp Med. 1994 May 1;179(5):1605–1611. doi: 10.1084/jem.179.5.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wang H., Liu K., Yuan F., Berdichevsky L., Taichman L. B., Auborn K. C/EBPbeta is a negative regulator of human papillomavirus type 11 in keratinocytes. J Virol. 1996 Jul;70(7):4839–4844. doi: 10.1128/jvi.70.7.4839-4844.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wolffe A. P. Structural and functional properties of the evolutionarily ancient Y-box family of nucleic acid binding proteins. Bioessays. 1994 Apr;16(4):245–251. doi: 10.1002/bies.950160407. [DOI] [PubMed] [Google Scholar]
  41. Wright K. L., Vilen B. J., Itoh-Lindstrom Y., Moore T. L., Li G., Criscitiello M., Cogswell P., Clarke J. B., Ting J. P. CCAAT box binding protein NF-Y facilitates in vivo recruitment of upstream DNA binding transcription factors. EMBO J. 1994 Sep 1;13(17):4042–4053. doi: 10.1002/j.1460-2075.1994.tb06721.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zachow K. R., Conklin K. F. CArG, CCAAT, and CCAAT-like protein binding sites in avian retrovirus long terminal repeat enhancers. J Virol. 1992 Apr;66(4):1959–1970. doi: 10.1128/jvi.66.4.1959-1970.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES