The long term objectives of this Program Project are to elucidate host and viral mechanisms that tilt the interaction of herpes simplex virus (HSV) and neurons either towards lytic infection or towards latency. HSV latency is the most fascinating biological property ofthe virus and its most important clinical feature, and understanding HSV latency may lead to new therapies or even a cure for this widespread pathogen. The three projects that are central to the Program Project all assess the effects of viral and host mutants or certain treatments on viral gene expression and chromatin status during establishment, maintenance, and reactivation of latency, either in mice or in vitro. The assays used for these measurements require specialized equipment, training, attention to detail, and careful analysis to guarantee that they are truly quantitative. Thus, to ensure standardization and quality and to allow comparisons of results across all three projects, it is highly desirable to have these assays performed by a core.
A specific aim of this core then is to isolate nucleic acids and perform standardized measurements of 1) viral DNA, which are normalized by measuring host DNA;2) viral mRNAs, which are normalized by measuring a host mRNA;3) viral miRNAs, which are normalized by measuring a host miRNA;and 4) occupancies by particular chromatin proteins on particular regions of viral DNA (chromatin immunoprecipitation, ChIP), which are normalized to occupancy on a host gene. These four assays use real-time polymerase chain reaction (PCR) based methods. An additional assay uses in situ hybridization to detect latency-associated transcripts in neurons.
A second aim of the core is to train personnel from the projects in these assays should there be too large a volume for the core or should the personnel wish to develop new assays.
A third aim i s to maintain the equipment needed to conduct the assays.

Public Health Relevance

The gene expression core performs important assays for the projects, and thus facilitates research to understand herpes simplex virus (HSV) latency, which is the most fascinating biological and most vexing clinical aspect of this virus. Understanding HSV latency might lead to new treatments or even cures for HSV infections, which are widespread and, in some cases, life-threatening.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI098681-02
Application #
8693917
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Harvard Medical School
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02115
Colgrove, Robert C; Liu, Xueqiao; Griffiths, Anthony et al. (2016) History and genomic sequence analysis of the herpes simplex virus 1 KOS and KOS1.1 sub-strains. Virology 487:215-21
Raja, Priya; Lee, Jennifer S; Pan, Dongli et al. (2016) A Herpesviral Lytic Protein Regulates the Structure of Latent Viral Chromatin. MBio 7:
Katzenell, Sarah; Leib, David A (2016) Herpes Simplex Virus and Interferon Signaling Induce Novel Autophagic Clusters in Sensory Neurons. J Virol 90:4706-19
Rosato, Pamela C; Katzenell, Sarah; Pesola, Jean M et al. (2016) Neuronal IFN signaling is dispensable for the establishment of HSV-1 latency. Virology 497:323-7
Lee, Jennifer S; Raja, Priya; Knipe, David M (2016) Herpesviral ICP0 Protein Promotes Two Waves of Heterochromatin Removal on an Early Viral Promoter during Lytic Infection. MBio 7:e02007-15
Rosato, Pamela C; Leib, David A (2015) Neuronal Interferon Signaling Is Required for Protection against Herpes Simplex Virus Replication and Pathogenesis. PLoS Pathog 11:e1005028
Rosato, Pamela C; Leib, David A (2015) Neurons versus herpes simplex virus: the innate immune interactions that contribute to a host-pathogen standoff. Future Virol 10:699-714
Knipe, David M (2015) Nuclear sensing of viral DNA, epigenetic regulation of herpes simplex virus infection, and innate immunity. Virology 479-480:153-9
Conwell, Sara E; White, Anne E; Harper, J Wade et al. (2015) Identification of TRIM27 as a novel degradation target of herpes simplex virus 1 ICP0. J Virol 89:220-9
Knipe, David M; Raja, Priya; Lee, Jennifer S (2015) Clues to mechanisms of herpesviral latent infection and potential cures. Proc Natl Acad Sci U S A 112:11993-4

Showing the most recent 10 out of 15 publications