First Advisor

Robert Millette

Term of Graduation

Spring 1997

Date of Publication

1997

Document Type

Thesis

Degree Name

Master of Science (M.S.) in Biology

Department

Biology

Language

English

Subjects

Myc oncogenes, Herpes simplex virus, Genetic regulation

DOI

10.15760/etd.8182

Physical Description

1 online resource (v, 99 pages)

Abstract

The purpose of this investigation was to identify the regulatory elements involved in the transactivation of the human c-myc gene by herpes simplex virus type-1 (HSV-1). This study mainly focused on a major c-myc promoter P2 (Fig. 1). Transient expression assays were carried out with a reporter plasmid, that contained either a wild type (+66wt c-myc) or mutant (+66 mt E2F, +66mt CT -I2) c-myc promoter fused to the chloramphenicol acetyltransferase (CAT) gene, cotransfected with effector plasmids that contained the HSV-1 immediate early (IE) genes ( encoding IE proteins, ICP4, ICP0, ICP27). The transient expression assays revealed that the HSV-1 IE protein, ICP4, was a major effector in transactivation of the human c-myc P2 promoter by HSV-1, and ICPO provided a synergistic effect with ICP4. Cis-acting elements E2F and CT-12, which were potential binding sites for cellular transcription factors E2F and Spl respectively, were shown to be important in HSV-1 induced c-myc transactivation. In addition, these assays indicated that ICPO influences ICP4 mediated activation through these two sites. Electrophoretic mobility shift assays, oligonucleotide competition assays, and footprinting analysis together demonstrated that viral proteins didn't directly interact with P2 promoter region. Three cellular transcription factors, Spl, E2F, and YYl, however, specifically interacted with the P2 promoter region, of which, Spl and its related proteins SpX bound to several other sites in addition of CT-I2 site.

Moreover, SpX also appeared to interact with another cellular factor that bound together with it on the P2 promoter region, and a second unknown cellular factor was also observed to interact with P2 promoter region.

Persistent Identifier

https://archives.pdx.edu/ds/psu/39768

Included in

Biology Commons

Share

COinS