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Principal Investigator: Daphne Christina Avgousti
Organization: FRED HUTCHINSON CANCER CENTER
Fiscal Year: 2024
Award: $484,000
Funding agency: National Institute of General Medical Sciences
PROJECT SUMMARY
The eukaryotic genome is compacted in a basic repeating structure called chromatin, the anomalous regulation
of which is characteristic of many diseases. In recent years, the technologies to study chromatin have leapt
forward, allowing us to investigate chromosome interactions in three dimensions. However, these methods rely
on measurements taken at a baseline state, thus, a barrier remains to understanding the biological functions of
chromatin in a dynamic system. It has become increasingly evident that viruses manipulate the nuclear
environment to generate viral progeny, necessarily hijacking host chromatin resources for viral benefit.
Therefore, virus infection provides an ideal dynamic system in which to investigate chromatin function. Moreover,
interrogating virus-host interactions has led to advances in virus biology and the discovery of some of the most
profound areas of molecular biology ranging from p53 to splicing. With the recent advances in chromatin
methodologies, we are now in an ideal position to take the next step in understanding chromosome biology in
the three-dimensional context of dynamic biological systems. In this proposal, we aim to employ cutting-edge
chromatin technology combined with virus infection to reveal fundamental chromatin functions. We
focus on nuclear replicating viruses, adenoviruses and multiple herpesviruses, as test cases for the virus to
pinpoint vulnerabilities in chromatin function exploited by pathogens. We have successfully identified three
scenarios in which cellular chromatin is distinctly reorganized for viral benefit: 1) adenovirus protein VII causes
global chromatin reorganization through linker histone displacement to disrupt the cell cycle and impact
transcription; 2) herpes simplex virus infection causes marginalization of host chromatin by generating new
regions of heterochromatin that promote egress of viral progeny; and 3) human cytomegalovirus polarizes
cellular chromatin to generate a functional viral assembly center. We will systematically investigate the
mechanisms by which these dramatic nuclear rearrangements occur in three dimensions using a combination of
high-resolution immunofluorescence microscopy, electron microscopy, and chromosome capture techniques. In
the previous funding period, our approaches defined multiple previously unknown vulnerabilities of chromatin,
positioning us to take the next step into understanding the mechanisms of chromatin organization in the nucleus
in 3D. Completion of these studies will identify new chromatin targets and assist development of innovative
therapies for cancer, inflammation, and viral diseases.
Terms: <3-D><3-Dimensional><3C-based approach><3C-based assay><3C-based method><3C-based strategy><3C-based technique><3C-based technology><3D><Adenoviridae><Adenovirus Protein><Adenoviruses><Antioncogene Protein p53><Area><Biological Function><Biological Process><Biology><CMV><Cancer Treatment><Cancers><Cell Body><Cell Cycle><Cell Division Cycle><Cell Nucleus><Cells><Cellular Tumor Antigen P53><Characteristics><Chromatin><Chromosomes><Cytomegalovirus><DNA Molecular Biology><Development><Disease><Disorder><Electron Microscopy><Environment><Funding><Gene Transcription><Genetic Transcription><Genome><HCMV><Herpes Simplex><Herpes Simplex Infections><Herpes simplex disease><Herpesviridae><Herpesvirus hominis disease><Herpesviruses><Heterochromatin><Histones><Human><Immunofluorescence Microscopy><Inflammation><Innovative Therapy><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Measurement><Methodology><Methods><Modern Man><Molecular Biology><Nuclear><Nucleus><Oncoprotein p53><P53><Phosphoprotein P53><Phosphoprotein pp53><Position><Positioning Attribute><Protein TP53><RNA Expression><RNA Splicing><Regulation><Research Resources><Resolution><Resources><Salivary Gland Viruses><Splicing><Structure><System><TP53><TP53 gene><TRP53><Techniques><Technology><Testing><Transcription><Tumor Protein p53><Tumor Protein p53 Gene><Viral><Viral Diseases><Virus><Virus Assembly><Virus Diseases><Virus Replication><anti-cancer therapy><biological systems><cancer therapy><cancer-directed therapy><chromatin conformation capture><chromosome capture><chromosome conformation capture><cytomegalovirus group><developmental><dynamic system><dynamical system><herpes simplex virus 1 infection><herpes simplex virus infection><herpes virus><malignancy><marginalization><neoplasm/cancer><p53 Antigen><p53 Genes><p53 Tumor Suppressor><pathogen><protein p53><resolutions><three dimensional><viral assembly><viral infection><viral multiplication><viral replication><virus host interaction><virus infection><virus multiplication><virus-induced disease>