Herpes simplex virus type 1 (HSV-1) is a widespread human pathogen with a seroprevalence of about 65% globally. The symptoms caused by an HSV-1 infection are most commonly mucosal lesions around the mouth (cold sores) and in rare cases inflammation of the brain (meningitis) or eyes (keratitis). Other medically relevant human pathogens can be found within the same virus family including varicella zoster virus, Epstein-Barr virus, cytomegalovirus and Kaposi sarcoma herpesvirus. Despite extensive research into herpesvirus biology, we still lack a complete understanding of the mechanism of herpesvirus genome replication. Genome replication is a fundamental process of any organism or virus. A profound understanding of the intricate mechanisms of viral genome replication will allow us to develop targeted antiviral drugs in the future.
We here revisited long-standing questions of the HSV-1 genome replication mechanism using cutting-edge single molecule imaging techniques. This approach enables the detection of transient structures and heterogeneous behaviour of individual molecules, leading to crucial insights into this fundamental process, which could not be gained using more conventional assays. The overall objective of this project was to gain a deeper understanding of herpesvirus replication mechanism by addressing unanswered questions in the field using latest imaging technology.
One of the objectives of this project was to assess the DNA structures, which are formed upon viral replication on the single-molecule level. We therefore first established an assay using TIRF microscopy to enable this. Another objective was to study the properties of the HSV-1 origin binding protein using novel imaging techniques.
