HSV1_Single_Molecule · Single molecule imaging of herpes simplex virus type 1 DNA replication
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2022-09-01 → 2024-08-31
- EU contribution
- €212,934
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Single molecule imaging of herpes simplex virus type 1 DNA replication
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.
Data: CORDIS, © European Union
Project objective
Herpes simplex virus type 1 (HSV-1) is a widespread and well-known pathogen. However, we still lack a complete understanding of the mechanism of HSV-1 genome replication. We propose to revisit 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 behavior, leading to further crucial insights into this fundamental process. The current model for HSV-1 genome replication comprises an origin-dependent followed by an origin-independent stage. Whereas we do have evidence and a good understanding of the origin-dependent phase, we completely lack evidence for the proposed structures of the origin-independent phase. Thus, in the proposed project we want to study aspects of the origin-dependent replication phase. Specifically, we want to reveal the role of the HSV-1 origin binding protein UL9 during replication as this protein was proposed to act as a switch from the first to the second stage. It has been shown that UL9 is an essential protein during the origin-dependent stage but dispensable and even inhibitory during the second, origin-independent stage. We here propose to use single-molecule imaging techniques to reveal the inhibitory effect of UL9 on HSV-1 mediated replication and to further characterize the specific conditions of UL9 binding to the origins of replication.
Original text from CORDIS.
Participants
- IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE · LondonCoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union