Human Cytomegalovirus (HCMV) is the major viral cause of birth defects, infecting 1-2% of all newborns annually. Approximately 5-10% of these congenitally infected infants will manifest signs of serious neurological damage at birth, which can include deafness, blindness, mental retardation and microcephaly. Another 10-15% will develop sensori-neural hearing loss and/or learning disabilities within the first 10 years of life. Our long-term goat is to understand the mechanism behind the development of morbidity and mortality in infants congenitally infected with HCMV. Over the last several years we have studied the interaction of HCMV with the cell cycle and DNA repair machinery of the permissively infected cell. We have discerned that HCMV sequesters many key regulatory and repair proteins into its viral replication centers. However, it appears to partition the components of several complexes so that all the proteins are present within the replication centers, but not all are available to the cellular genome. We have also determined that HCMV can induce specific damage on chromosome 1 during S-phase. When coupled with the literature regarding nonspecific damage induced at late times post infection our data more clearly highlights the genotoxic effects of HCMV. We hypothesize that long-term detrimental consequences to the cellular genome may occur if 1) the initial specific damage is propagated or 2) damage incurred at late times post infection is not repaired due to sequestration of the repair machinery. To test our hypothesis, we propose three specific aims. First, we will thoroughly define the parameters of chromosome lq breakage in HCMV-infected cells with regard to rapidity of induction and cell cycle phase at time of infection. We think it is imperative that our results be moved into more clinically relevant cell types, especially cells of neural lineage, as these are the cells most severely affected by the virus during congenital infection. Second, we also will determine the consequences of chromosome 1 damage in these clinically relevant cells, and whether in a semi-permissive environment we can observe propagation of the chromosome 1 damage instead of healing of the break or movement of the cell toward apoptosis. Lastly, we will characterize the ability of HCMV-infected cells to repair exogenously introduced damage at late times post infection, after viral replication centers are assembled.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI051463-05
Application #
7155531
Study Section
Virology Study Section (VR)
Program Officer
Beisel, Christopher E
Project Start
2002-12-15
Project End
2008-11-30
Budget Start
2006-12-01
Budget End
2008-11-30
Support Year
5
Fiscal Year
2007
Total Cost
$244,523
Indirect Cost
Name
University of Idaho
Department
Microbiology/Immun/Virology
Type
Schools of Earth Sciences/Natur
DUNS #
075746271
City
Moscow
State
ID
Country
United States
Zip Code
83844
Kuan, Man I; O'Dowd, John M; Fortunato, Elizabeth A (2016) The absence of p53 during Human Cytomegalovirus infection leads to decreased UL53 expression, disrupting UL50 localization to the inner nuclear membrane, and thereby inhibiting capsid nuclear egress. Virology 497:262-278
Zavala, Anamaria G; O'Dowd, John M; Fortunato, Elizabeth A (2016) Infection of a Single Cell Line with Distinct Strains of Human Cytomegalovirus Can Result in Large Variations in Virion Production and Facilitate Efficient Screening of Virus Protein Function. J Virol 90:2523-35
Kuan, Man I; O'Dowd, John M; Chughtai, Kamila et al. (2016) Human Cytomegalovirus nuclear egress and secondary envelopment are negatively affected in the absence of cellular p53. Virology 497:279-293
Fortunato, Elizabeth A (2014) Use of diploid human fibroblasts as a model system to culture, grow, and study human cytomegalovirus infection. Methods Mol Biol 1119:47-57
Kulkarni, Amit S; Fortunato, Elizabeth A (2014) Modulation of homology-directed repair in T98G glioblastoma cells due to interactions between wildtype p53, Rad51 and HCMV IE1-72. Viruses 6:968-85
Duan, Ying-Liang; Ye, Han-Qing; Zavala, Anamaria G et al. (2014) Maintenance of large numbers of virus genomes in human cytomegalovirus-infected T98G glioblastoma cells. J Virol 88:3861-73
Zavala, Anamaria G; Kulkarni, Amit S; Fortunato, Elizabeth A (2014) A dual color Southern blot to visualize two genomes or genic regions simultaneously. J Virol Methods 198:64-8
Dziurzynski, Kristine; Chang, Susan M; Heimberger, Amy B et al. (2012) Consensus on the role of human cytomegalovirus in glioblastoma. Neuro Oncol 14:246-55
Duan, Yingliang; Miao, Lingfeng; Ye, Hanqing et al. (2012) A faster immunofluorescence assay for tracking infection progress of human cytomegalovirus. Acta Biochim Biophys Sin (Shanghai) 44:597-605
O'Dowd, John M; Zavala, Anamaria G; Brown, Celeste J et al. (2012) HCMV-infected cells maintain efficient nucleotide excision repair of the viral genome while abrogating repair of the host genome. PLoS Pathog 8:e1003038

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