Multidisciplinary molecular genetics and biochemical approaches are being used to study replication of avian and human oncogenic retroviruses. Areas of particular interest are in reverse transcription, viral DNA integration, and virion assembly.
Specific projects include studying
1) the role of viral RNA secondary structures in initiation of reverse transcription,
2) the mechanism of concerted integration of viral DNA into the host chromosome by integrase (IN) using an in vitro reconstituted system,
3) the mechanism of processing of Gag and Pol polyproteins into mature viral proteins by the virus-specific protease (PR), and
4) the role of a Gag polyprotein assembly domain in budding of virus particles from cells.
In many of these studies, amino acid substitutions have been placed at biochemically or structurally important residues and the effect these changes have on viral replication and on the properties of the mutant proteins have been defined.
Selected Publications:
Brin, E., Jizu, Y, Skalka, A.M., & Leis, J. Modeling the late steps in HIV-1 IN-catalyzed DNA integration. J. Biol. Chem. 275:39287-39295 (2000).
VerPlank, L., Agresta, B., Grassa, T., Kikonyogo, A., Leis, J., & Carter, C. Tsg101, the prototype of a class of dominant-negative ubiquitin regulators, binds human immunodeficiency virus type 1 Pr55Gag: the L domain is a determining of binding. Proc. Natl. Acad. Sci. USA 98:7724-7729 (2001). PMID: 11427703
Kikonyogo, A., Bouamr, F., Vana, M., Xiang, Y., Aiyar, A., Carter, C., and Leis, J. Proteins related to the Nedd4 family of ubiquitin protein ligases interact with the L domain of Rous sarcoma virus and are required for Gag budding from cells. Proc. Natl. Acad. Sci. USA. . 98: 11199-11204 (2001). PMID: 11562473
Vana, M. L., Tang, Y., Chen, A., Medina, G., Carter, C., and Leis, J. The role of Nedd4 and ubiquitination of RSV Gag in budding of virus-like particles from cells. J. Virol. 78, 13943-13953 (2004). PMID: 15564502
Medina, G., Pincetic, A., Zhang, Y., Ehrlich, L. S., Tang, Y., Leis, J., and Carter, C. Tsg101 can replace Nedd4 function in ASV Gag release but not membrane targeting. Virology 377: 30-38 (2008). PMID: 18555885
Pincetic, A., Medina, G., Carter, C., and Leis, J. Avian Sarcoma Virus and Human Immunodeficiency Virus, Type 1 use Different Subsets of ESCRT Proteins to Facilitate the Budding Process. J. Biol. Chem. 283, 29822-29830 (2008). PMID: 18723511
Dolan, J., Chen, A., Weber, I.T, Harrison, R.W., and Leis, J. Defining the DNA Substrate Binding Sites on HIV-1 Integrase. JMB 385, 568-579 (2009). PMID: 19014951
