VACCINIA VIRUS O1L VIRULENCE GENE AND PROTEIN LOCALIZATION

Abstract

Smallpox killed an estimated 500 million people in the twentieth century alone. Although this fatal disease was eradicated from the world over thirty years ago, its potential use as a bioterrorism agent remains a concern. In addition, monkeypox continues to cause human outbreaks in Africa, and in the US in 2003. Vaccinia virus, the live virus vaccine for smallpox and monkeypox, is dangerous for immunocompromised individuals, and a safer vaccine is needed. The Roper lab studies how poxviruses cause disease in mammals and which genes contribute to virulence. The vaccinia virus O1L gene is highly conserved in poxviruses, and we have shown that it is required for full virulence in mice. When the O1L gene is removed from the wild type virus, the virus becomes attenuated, and immune responses are improved. Very little is known about this protein including its molecular weight, location within the cell and its function. We raised anti O1L peptide antibodies in rabbits and are using these to investigate the localization of the O1L protein using immunofluorescence techniques. In accordance with preliminary data from western blot analysis, we hypothesized that the O1L protein is located in the nucleus of the cell. Through immunofluorescence, the O1L protein was detected in the nucleus and cytoplasm of the cell. Identifying where the protein localizes will help us to form further hypotheses as to the mechanism of O1L immunosuppression and understand how poxviruses control the immune system. This research will increase our understanding of poxvirus pathogenesis and aid in the creation of new and safer vaccines.