Project B2 - Viral macrodomain proteins as therapeutic targets for coronavirus infections
Prof. Dr. med. John Ziebuhr
Department of Medical Virology
Biomedical Research Center Seltersberg (BFS)
Justus Liebig University Giessen
Tel.: +49 641 99 41200
Fax: +49 641 99 41209
- Project description
Background: Coronaviruses encode enzymes that other RNA viruses do not (or rarely) encode. These include, for example, exo- and endoribonucleases, methyltransferases, 2'-5' phosphodiesterases, and macrodomain proteins 1,2,3,4,5,6. For coronaviral macrodomains, specific functions in viral pathogenesis and replication efficiency have been suggested7. Other medically important viruses such as chikungunya and hepatitis E were also found to encode macrodomains, suggesting that these domains represent suitable targets for the development of broad-spectrum antivirals against viruses from different families. Previous work in the laboratory led to the identification of coronavirus macrodomain-associated ADP-ribose-1"-phosphatase activities (SARS-CoV, HCoV-229E, TGEV) 8,9,10,11. There is also evidence that macrodomain proteins can bind to polyADP-ribose and hydrolyze mono-ADP-ribosylated substrates10,12.
Scientific goals: Based on previous work, the substrate specificities of viral macrodomains will be characterized in more detail, suitable HTS assays will be developed, and possible effects of coronaviral macrodomain activities on viral replication in different types of host cells will be determined: (1) expression and purification of corona- and alphavirus macrodomain proteins and biochemical characterization; (2) generation and characterization of coronavirus macrodomain knock-out mutants; (3) development of HTS assays; (4) comparative proteome and transcriptome analyses of virus-infected cells (wild type versus mutants); and (5) validation of selected proteome and transcriptome data.
References B2: 1. Minskaia et al. (2006) Proc Natl Acad Sci USA 103:5108-5113.* 2. Snijder et al. (2016) Adv Virus Res 96:59-126.* 3. Kindler et al. (2017) PLoS Pathog 13:e1006195 4. Züst et al. (2011) Nature Immunol 12:137-143.* 5. Habjan et al. (2013) PLoS Pathogens 9:e1003663.* 6. Zhao et al. (2012) Cell Host Microbe 11:607-616.* 7. Eriksson et al. (2008) J Virol 82:12325-12334. 8. Putics et al. (2005) J Virol 79:12721-12731.* 9. Putics et al. (2006) J Gen Virol 87:651-656.* 10. Egloff et al. (2006) J Virol 80:8493-8502.* 11. Kuri et al. (2011) J Gen Virol 92:1899-1905.* 12. Li et al. (2016) J Virol 90:8478-8486.
* author’s own publications