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.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
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)
Barklis, E., Mulligan, R. C. & Jaenisch, R. Chromosomal position or virus mutation permits retrovirus expression in embryonal carcinoma cells. Cell 47, 391–399 (1986)
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)
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)
Yamauchi, M. et al. Stem cell factor binding to retrovirus primer binding site silencers. J. Virol. 69, 1142–1149 (1995)
Wolf, D. & Goff, S. P. Host restriction factors blocking retroviral replication. Annu. Rev. Genet. 42, 143–163 (2008)
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)
Wolf, D. & Goff, S. P. TRIM28 mediates primer binding site-targeted silencing of murine leukemia virus in embryonic cells. Cell 131, 46–57 (2007)
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)
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)
O’Geen, H. et al. Genome-wide analysis of KAP1 binding suggests autoregulation of KRAB-ZNFs. PLoS Genet. 3, e89 (2007)
Tsuruma, R. et al. Physical and functional interactions between STAT3 and KAP1. Oncogene 27, 3054–3059 (2008)
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)
Berg, J. M. & Shi, Y. The galvanization of biology: a growing appreciation for the roles of zinc. Science 271, 1081–1085 (1996)
Urrutia, R. KRAB-containing zinc-finger repressor proteins. Genome Biol. 4, 231 (2003)
Friedman, J. R. et al. KAP-1, a novel corepressor for the highly conserved KRAB repression domain. Genes Dev. 10, 2067–2078 (1996)
Fuks, F. et al. The methyl-CpG-binding protein MeCP2 links DNA methylation to histone methylation. J. Biol. Chem. 278, 4035–4040 (2003)
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)
Bex, F. & Gaynor, R. B. Regulation of gene expression by HTLV-I Tax protein. Methods 16, 83–94 (1998)
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)
Wolf, D. et al. Acetylation of β-catenin by CREB-binding protein (CBP). J. Biol. Chem. 277, 25562–25567 (2002)
Telesnitsky, A., Blain, S. & Goff, S. P. Assays for retroviral reverse transcriptase. Methods Enzymol. 262, 347–362 (1995)
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.
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
The Regulation and Functions of Endogenous Retrovirus in Embryo Development and Stem Cell Differentiation
Stem Cells International (2021)
Argonaute NRDE-3 and MBT domain protein LIN-61 redundantly recruit an H3K9me3 HMT to prevent embryonic lethality and transposon expression
Genes & Development (2021)
The FEBS Journal (2021)
Frontiers in Cellular and Infection Microbiology (2021)
Conserved paradoxical relationships among the evolutionary, structural and expressional features of KRAB zinc-finger proteins reveal their special functional characteristics
BMC Molecular and Cell Biology (2021)