Virus Genes

, Volume 45, Issue 2, pp 265–273 | Cite as

CTCF and Sp1 interact with the Murine gammaherpesvirus 68 internal repeat elements

  • Hannah C. Stevens
  • Kevin S-W Cham
  • David J. Hughes
  • Ren Sun
  • Jeffery T. Sample
  • Vivien J. Bubb
  • James P. Stewart
  • John P. Quinn


Herpesviruses maintain a dynamic balance between latency and productive infection. This is a complex process regulated by viral and cellular factors. We have developed a Murine gammaherpesvirus 68 (MHV-68) model system in which to study mechanisms underlying balance between latency and lytic infection. We have generated an epithelial cell line that carries MHV-68 in a tightly latent form by using a bacterial artificial chromosome clone of the virus genome with a mutation in the MHV-68 major lytic R transactivator gene. Complementation of this defect in trans by transfection with a plasmid encoding R transactivator initiated and restored the productive cycle. This cell line model was used to investigate transcription factor occupancy (CCCTC binding factor [CTCF] and Sp1) of the two internal repeat elements in the viral genome during latency and reactivation using chromatin immunoprecipitation. Our results show that CTCF can bind to the 40-bp and the 100-bp repeat sequences during latency, whereas binding is reduced upon reactivation. In contrast, Sp1 only bound to the 100-bp repeat after reactivation. Our results indicate that the large internal repeat sequences in MHV-68 have different functions. We hypothesise that the 40-bp repeat may be involved in regulation of gene expression during the maintenance of latency, while the 100-bp repeat domain may be involved in regulation of the lytic cycle.


Herpesvirus Latency Reactivation Transcriptional control Mouse model CCCTC binding factor Sp1 



HCS was funded by the Wellcome Trust Prize Studentship. JPQ and VJB were funded by the BBSRC (award BB/D016754/9). This work was funded in part by U.S. Public Health Service grant CA090208 from the National Cancer Institute and the Penn State Hershey Cancer Institute. JPS was funded by a Royal Society (London) University Research Fellowship. The authors would like to thank Dr Bahram Ebrahimi for the kind gift of pFLAG-CMV1-ORF50.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Hannah C. Stevens
    • 1
  • Kevin S-W Cham
    • 2
  • David J. Hughes
    • 2
    • 5
  • Ren Sun
    • 3
  • Jeffery T. Sample
    • 4
  • Vivien J. Bubb
    • 1
  • James P. Stewart
    • 2
  • John P. Quinn
    • 1
  1. 1.Department of Molecular and Clinical PharmacologyInstitute of Translational Medicine, University of LiverpoolLiverpoolUK
  2. 2.Department of Infection BiologyUniversity of LiverpoolLiverpoolUK
  3. 3.Department of Molecular and Medical PharmacologyUniversity of California at Los AngelesLos AngelesUSA
  4. 4.Department of Microbiology and ImmunologyThe Pennsylvania State University College of MedicineHersheyUSA
  5. 5.Institute of Molecular and Cellular Biology, Faculty of Biological SciencesUniversity of LeedsLeedsUK

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