Human cytomegalovirus (HCMV) is a ubiquitous herpes virus causing mild or subclinical disease in immunocompetent adults. However, severe complications may be encountered in immunocompromised individuals. HCMV is a systemic infection that may infect several sites in the body, including the retina, gastrointestinal tract, lungs, liver and central nervous system. Initially it is was shown that human cytomegalovirus (HCMV) required eleven distinct loci for origin-dependent DNA replication. In later studies it was demonstrated that, of the eleven loci originally described, UL84 appeared to be the only non- core replication protein required for oriLyt-dependent replication. This fact, coupled with the observation that RNA-DNA hybrid structures are present within HCMV oriLyt, suggests the possibility that HCMV may have a mode of initiation of DNA replication unlike that of other herpesviruses. UL84 may be the key factor involved in initiation of HCMV DNA replication and, therefore, an ideal target for chemotherapeutic agents needed to control infection. To this end, this proposal will define the role of UL84 in the initiation of HCMV DNA replication and effects on cellular processes. The UL84 protein is required for HCMV DNA replication as demonstrated in transient assays and with a viral mutant. The elucidation of a function for UL84 will allow for the development of anti-HCMV drugs. Our hypothesis is that UL84 is involved in the initiation of HCMV DNA replication by a direct interaction with the origin of replication, oriLyt, and participates in the regulation of host cell RNA processing.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI045096-02
Application #
6341734
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Beisel, Christopher E
Project Start
2000-01-15
Project End
2004-12-31
Budget Start
2001-01-01
Budget End
2001-12-31
Support Year
2
Fiscal Year
2001
Total Cost
$271,233
Indirect Cost
Name
University of Nevada Reno
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
146515460
City
Reno
State
NV
Country
United States
Zip Code
89557
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Gao, Yang; Kagele, Dominique; Smallenberg, Kate et al. (2010) Nucleocytoplasmic shuttling of human cytomegalovirus UL84 is essential for virus growth. J Virol 84:8484-94
Kagele, Dominique; Gao, Yang; Smallenburg, Kate et al. (2009) Interaction of HCMV UL84 with C/EBPalpha transcription factor binding sites within oriLyt is essential for lytic DNA replication. Virology 392:16-23
Gao, Yang; Pari, Gregory S (2009) Interaction of human cytomegalovirus pUL84 with casein kinase 2 is required for oriLyt-dependent DNA replication. J Virol 83:2393-6
Gao, Yang; Colletti, Kelly; Pari, Gregory S (2008) Identification of human cytomegalovirus UL84 virus- and cell-encoded binding partners by using proteomics analysis. J Virol 82:96-104
Colletti, Kelly S; Smallenburg, Kate E; Xu, Yiyang et al. (2007) Human cytomegalovirus UL84 interacts with an RNA stem-loop sequence found within the RNA/DNA hybrid region of oriLyt. J Virol 81:7077-85
Colletti, Kelly S; Xu, Yiyang; Yamboliev, Irena et al. (2005) Human cytomegalovirus UL84 is a phosphoprotein that exhibits UTPase activity and is a putative member of the DExD/H box family of proteins. J Biol Chem 280:11955-60
Xu, Yiyang; Cei, Sylvia A; Rodriguez Huete, Alicia et al. (2004) Human cytomegalovirus DNA replication requires transcriptional activation via an IE2- and UL84-responsive bidirectional promoter element within oriLyt. J Virol 78:11664-77
Colletti, Kelly S; Xu, Yiyang; Cei, Sylvia A et al. (2004) Human cytomegalovirus UL84 oligomerization and heterodimerization domains act as transdominant inhibitors of oriLyt-dependent DNA replication: evidence that IE2-UL84 and UL84-UL84 interactions are required for lytic DNA replication. J Virol 78:9203-14

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