Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Embryonic stem cells use ZFP809 to silence retroviral DNAs

Abstract

Embryonic stem cells (ESCs) and other primitive stem cells of mice have been known for more than 30 years to potently block retrovirus replication1. Infection of ESCs by the murine leukaemia viruses (MLVs) results in the normal establishment of integrated proviral DNA, but this DNA is then transcriptionally silenced, preventing further viral spread. The repression is largely mediated by trans-acting factors that recognize a conserved sequence element termed the primer binding site, an 18-base pair sequence complementary to the 3′ end of a cellular transfer RNA2,3,4,5,6. A specific tRNA is annealed to the primer binding site sequence of the viral genomic RNA, and is used to prime DNA synthesis7. This same sequence in the context of the integrated proviral DNA is targeted for silencing in ESCs. We have recently shown that a large protein complex binding to the primer binding site in ESCs contains TRIM28 (refs 8, 9), a well-characterized transcriptional co-repressor10,11,12. An important question remains as to the identity of the factor that directly recognizes integrated retroviral DNAs and recruits TRIM28 to mediate their specific silencing. Here we identify the zinc finger protein ZFP809 as the recognition molecule that bridges the integrated proviral DNA and TRIM28. We show that expression of ZFP809 is sufficient to render even differentiated cells highly resistant to MLV infection. Furthermore, we demonstrate that ZFP809 is able to potently block transcription from DNA constructs of human T-cell lymphotropic virus-1 (HTLV-1), which use the same primer tRNA. These results identify ZFP809 as a DNA-binding factor that specifically recognizes a large subset of mammalian retroviruses and retroelements, targeting them for transcriptional silencing. We propose that ZFP809 evolved as a stem-cell-specific retroviral restriction factor, and therefore constitutes a new component of the intrinsic immune system of stem cells.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Identification of candidate PBS Pro DNA-binding factors.
Figure 2: Reconstitution of the PBS Pro binding complex with recombinant proteins.
Figure 3: Stable expression of ZFP809 in 293A cells leads to reconstitution of the PBS silencing complex in vivo.
Figure 4: Expression of ZFP809 in a differentiated cell line causes a potent block to the replication of PBS Pro -using retroviruses.

References

  1. 1

    Teich, N. M., Weiss, R. A., Martin, G. R. & Lowy, D. R. Virus infection of murine teratocarcinoma stem cell lines. Cell 12, 973–982 (1977)

    CAS  Article  Google Scholar 

  2. 2

    Barklis, E., Mulligan, R. C. & Jaenisch, R. Chromosomal position or virus mutation permits retrovirus expression in embryonal carcinoma cells. Cell 47, 391–399 (1986)

    CAS  Article  Google Scholar 

  3. 3

    Feuer, G., Taketo, M., Hanecak, R. C. & Fan, H. Two blocks in Moloney murine leukemia virus expression in undifferentiated F9 embryonal carcinoma cells as determined by transient expression assays. J. Virol. 63, 2317–2324 (1989)

    CAS  PubMed  PubMed Central  Google Scholar 

  4. 4

    Petersen, R., Kempler, G. & Barklis, E. A stem cell-specific silencer in the primer-binding site of a retrovirus. Mol. Cell. Biol. 11, 1214–1221 (1991)

    CAS  Article  Google Scholar 

  5. 5

    Yamauchi, M. et al. Stem cell factor binding to retrovirus primer binding site silencers. J. Virol. 69, 1142–1149 (1995)

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 6

    Wolf, D. & Goff, S. P. Host restriction factors blocking retroviral replication. Annu. Rev. Genet. 42, 143–163 (2008)

    CAS  Article  Google Scholar 

  7. 7

    Harada, F., Peters, G. G. & Dahlberg, J. E. The primer tRNA for Moloney murine leukemia virus DNA synthesis. Nucleotide sequence and aminoacylation of tRNAPro . J. Biol. Chem. 254, 10979–10985 (1979)

    CAS  PubMed  Google Scholar 

  8. 8

    Wolf, D. & Goff, S. P. TRIM28 mediates primer binding site-targeted silencing of murine leukemia virus in embryonic cells. Cell 131, 46–57 (2007)

    CAS  Article  Google Scholar 

  9. 9

    Wolf, D., Cammas, F., Losson, R. & Goff, S. P. Primer binding site-dependent restriction of murine leukemia virus requires HP1 binding by TRIM28. J. Virol. 82, 4675–4679 (2008)

    CAS  Article  Google Scholar 

  10. 10

    Ayyanathan, K. et al. Regulated recruitment of HP1 to a euchromatic gene induces mitotically heritable, epigenetic gene silencing: a mammalian cell culture model of gene variegation. Genes Dev. 17, 1855–1869 (2003)

    CAS  Article  Google Scholar 

  11. 11

    O’Geen, H. et al. Genome-wide analysis of KAP1 binding suggests autoregulation of KRAB-ZNFs. PLoS Genet. 3, e89 (2007)

    Article  Google Scholar 

  12. 12

    Tsuruma, R. et al. Physical and functional interactions between STAT3 and KAP1. Oncogene 27, 3054–3059 (2008)

    CAS  Article  Google Scholar 

  13. 13

    Kolosha, V. O. & Martin, S. L. In vitro properties of the first ORF protein from mouse LINE-1 support its role in ribonucleoprotein particle formation during retrotransposition. Proc. Natl Acad. Sci. USA 94, 10155–10160 (1997)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Berg, J. M. & Shi, Y. The galvanization of biology: a growing appreciation for the roles of zinc. Science 271, 1081–1085 (1996)

    ADS  CAS  Article  Google Scholar 

  15. 15

    Urrutia, R. KRAB-containing zinc-finger repressor proteins. Genome Biol. 4, 231 (2003)

    Article  Google Scholar 

  16. 16

    Friedman, J. R. et al. KAP-1, a novel corepressor for the highly conserved KRAB repression domain. Genes Dev. 10, 2067–2078 (1996)

    CAS  Article  Google Scholar 

  17. 17

    Fuks, F. et al. The methyl-CpG-binding protein MeCP2 links DNA methylation to histone methylation. J. Biol. Chem. 278, 4035–4040 (2003)

    CAS  Article  Google Scholar 

  18. 18

    Felber, B. K. et al. The pX protein of HTLV-I is a transcriptional activator of its long terminal repeats. Science 229, 675–679 (1985)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Bex, F. & Gaynor, R. B. Regulation of gene expression by HTLV-I Tax protein. Methods 16, 83–94 (1998)

    CAS  Article  Google Scholar 

  20. 20

    Wolf, D., Hug, K. & Goff, S. P. TRIM28 mediates primer binding site-targeted silencing of Lys1,2 tRNA-utilizing retroviruses in embryonic cells. Proc. Natl Acad. Sci. USA 105, 12521–12526 (2008)

    ADS  CAS  Article  Google Scholar 

  21. 21

    Wolf, D. et al. Acetylation of β-catenin by CREB-binding protein (CBP). J. Biol. Chem. 277, 25562–25567 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Telesnitsky, A., Blain, S. & Goff, S. P. Assays for retroviral reverse transcriptase. Methods Enzymol. 262, 347–362 (1995)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This work was supported by Public Health Service grant R37 CA 30488 from the National Cancer Institute. We thank D. Derse, F. Bouamr and E. Barklis for their generosity with reagents. We are grateful to M. de los Santos, H. Nickerson and M. A. Gawinowicz (Protein Core Facility Columbia University) for technical assistance. We would also like to thank P. Adusumilli, M. Banes, M. Bilsky, J. Yamada and co-workers, without whose help this paper would never have been written. D.W. is an Associate, and S.P.G. is an Investigator of the Howard Hughes Medical Institute.

Author Contributions D.W. performed the experiments and analysed the data. D.W. and S.P.G. designed the study and wrote the manuscript.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Stephen P. Goff.

Supplementary information

Supplementary Information

This file contains a Supplementary Table, Supplementary Figures 1-8 with Legends and a Supplementary Reference. (PDF 1243 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wolf, D., Goff, S. Embryonic stem cells use ZFP809 to silence retroviral DNAs. Nature 458, 1201–1204 (2009). https://doi.org/10.1038/nature07844

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing