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Start of funding 01.01.2009

Recombinant herpesviruses for research in autophagy and cancer

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The project will combine the Erlangen recombinant DNA technology know-how, specifically the genetic manipulation of herpesviral genomes by homologous recombination, with the USC platforms in cell biology and immunology. It is planned to create recombinant KSHV viruses with specific marker gene insertion and gene deletion relevant to virus induced signal transduction, tumorigenesis and autophagy. These viruses will then be analyzed by the state-of-the-art cell biology and immunology assays established in Prof. Jung’s laboratory at USC. The techniques and results that are derived from this cooperation not only will be used for a joint research program focused on antiviral and antitumor host defence, but also will ultimately lead to the discovery of new approaches in the therapy of virus associated human cancer.

Final report:
The BaCaTeC project funding initiated a fruitful cooperation between Californian and Bavarian laboratories. The Kaposi's sarcoma herpesvirus (KSHV) bacterial artificial chromosome (BACmid ) Bac16, newly produced at USC in the laboratory of Prof. Jung, was transferred by us into appropriate host bacteria and can now be changed rapidly and specifically by homologous recombination. This KSHV Bac16 platform (Brulois 2012) was also successfully established in Erlangen, where we now are also to produce recombinant KSHV. In conjunction with a specific inducible host cell line this system represents the state of the art for the virus-genetic analysis of KSHV. As an example, we could now demonstrate a function of the viral K4.2 protein as a regulator of immunoglobulin secretion and calcium homeostasis in the endoplasmic reticulum (Wong 2013). Finally, we were just able to uncover a new function of the viral effector ORF75, a viral tegument protein, in the modulation of intrinsic cellular restriction factors in nuclear domain 10 (ND10) structures (Full 2013).
In the future, our U.S. American and German laboratories will continue this successful cooperation on the pathogenesis of Kaposi's sarcoma and also on KSHV-associated hematologic neoplasias. The success of the collaboration is already documented by relevant publications in leading infectiological journals such as the Journal of Virology and PLOS Pathogens.

Publications

Brulois K, Chang H, Lee A, Ensser A, Wong LY, Toth Z, Lee SH, Lee H-R, Myoung J, Ganem D, Oh T-K, Kim JF, Gao SJ, Jung JU (2012) Construction and manipulation of a new Kaposi’s sarcoma-associated herpesvirus Bacterial artificial chromosome clone. J Virol. 86: 9708-9720.

Wong LY, Brulois K, Toth Z, Inn KS, Lee SH, O'Brien K, Lee H, Gao SJ, Cesarman E, Ensser A, Jung JU (2013) The Product of Kaposi's Sarcoma-Associated Herpesvirus Immediate Early Gene K4.2 Regulates Immunoglobulin Secretion and Calcium Homeostasis by Interacting with and Inhibiting pERP1. J Virol. 87:12069-79.

Full F, Lengenfelder D, Reuter N, Bogner E, Brulois K, Scholz B, Stürzl M, Myoung J, Jung JU, Stamminger T, Ensser A (2013) Kaposi's sarcoma associated herpesvirus tegument protein ORF75 is essential for viral lytic replication and plays a critical role in the antagonization of ND10-instituted intrinsic immunity. PLOS Pathogens, in press.