Human cytomegalovirus (HCMV), a member of the herpesvirus family, is an important pathogen capable of causing a variety of diseases. HCMV is the leading viral cause of birth defects and poses a serious health threat to immunocompromised individuals, particularly those with AIDS. The multiple pathogenic effects of HCMV are likely manifested through a complex interplay of viral gene products and induced and repressed cellular functions. The research in Dr. Spector's laboratory has been directed towards characterizing the sequence requirements and trans- acting factors (both viral and cellular) necessary for the control of three representative early transcription units including: the 2.7 kb and 1.2 kb RNA transcripts encoded by the repeat bounding the long unique segment of the genome; and the 2.2 kb family of RNA transcripts encoding 4 related nuclear phosphoproteins. In this renewal application, the investigator proposes to continue their studies on the regulation of HCMV early gene expression and the functional properties of the major viral transactivator, an immediate early protein designated IE2 86. The approach is to couple functional in vivo genetic analyses with in vitro biochemical and molecular assays to achieve the following aims: (1) determination of the cis-acting regulatory elements on HCMV early promoters when the promoters are part of the infectious viral genome; (2) analysis of the function of the IE2 86 protein in the context of mutant recombinant viruses; (3) identification and characterization of the trans-acting cellular factors interacting with the HCMV 1.2 kb and 2.2 kb early RNA promoters; (4) use of in vitro RNA pol I transcription reactions to assess the functional basis of interaction of the IE2 86 protein with the upstream binding factor UBF; and (5) utilization of in vitro RNA pol II transcription reactions to determine the molecular mechanisms by which the IE2 86 protein stimulates HCMV early promoters. The long range goals of this research are to define at the molecular level the mechanisms which operate to control HCMV gene expression, thus providing an important foundation for understanding how the complex interactions of viral and host functions relate to the pathogenesis of the virus.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA034729-14
Application #
2429645
Study Section
Virology Study Section (VR)
Project Start
1983-05-01
Project End
2001-05-31
Budget Start
1997-06-01
Budget End
1998-05-31
Support Year
14
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Clark, Elizabeth; Spector, Deborah H (2015) Studies on the Contribution of Human Cytomegalovirus UL21a and UL97 to Viral Growth and Inactivation of the Anaphase-Promoting Complex/Cyclosome (APC/C) E3 Ubiquitin Ligase Reveal a Unique Cellular Mechanism for Downmodulation of the APC/C Subunits APC1, J Virol 89:6928-39
Strang, Blair L; Bender, Brian J; Sharma, Mayuri et al. (2012) A mutation deleting sequences encoding the amino terminus of human cytomegalovirus UL84 impairs interaction with UL44 and capsid localization. J Virol 86:11066-77
DuRose, Jenny B; Li, Julie; Chien, Shu et al. (2012) Infection of vascular endothelial cells with human cytomegalovirus under fluid shear stress reveals preferential entry and spread of virus in flow conditions simulating atheroprone regions of the artery. J Virol 86:13745-55
Burgdorf, Sarah W; Clark, Charles L; Burgdorf, James R et al. (2011) Mutation of glutamine to arginine at position 548 of IE2 86 in human cytomegalovirus leads to decreased expression of IE2 40, IE2 60, UL83, and UL84 and increased transcription of US8-9 and US29-32. J Virol 85:11098-110
Tran, Karen; Mahr, Jeffrey A; Spector, Deborah H (2010) Proteasome subunits relocalize during human cytomegalovirus infection, and proteasome activity is necessary for efficient viral gene transcription. J Virol 84:3079-93
Tran, Karen; Kamil, Jeremy P; Coen, Donald M et al. (2010) Inactivation and disassembly of the anaphase-promoting complex during human cytomegalovirus infection is associated with degradation of the APC5 and APC4 subunits and does not require UL97-mediated phosphorylation of Cdh1. J Virol 84:10832-43
Sanders, Rebecca L; Spector, Deborah H (2010) Human cytomegalovirus IE2 86 and IE2 40 proteins differentially regulate UL84 protein expression posttranscriptionally in the absence of other viral gene products. J Virol 84:5158-70
Kapasi, Anokhi J; Clark, Charles L; Tran, Karen et al. (2009) Recruitment of cdk9 to the immediate-early viral transcriptosomes during human cytomegalovirus infection requires efficient binding to cyclin T1, a threshold level of IE2 86, and active transcription. J Virol 83:5904-17
Kapasi, Anokhi J; Spector, Deborah H (2008) Inhibition of the cyclin-dependent kinases at the beginning of human cytomegalovirus infection specifically alters the levels and localization of the RNA polymerase II carboxyl-terminal domain kinases cdk9 and cdk7 at the viral transcriptosome. J Virol 82:394-407
Sanders, Rebecca L; Clark, Charles L; Morello, Christopher S et al. (2008) Development of cell lines that provide tightly controlled temporal translation of the human cytomegalovirus IE2 proteins for complementation and functional analyses of growth-impaired and nonviable IE2 mutant viruses. J Virol 82:7059-77

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