First Advisor

Robert L. Millette

Term of Graduation

Summer 1994

Date of Publication

7-6-1994

Document Type

Thesis

Degree Name

Master of Science (M.S.) in Biology

Department

Biology

Language

English

Subjects

Nucleotide sequence, Herpes simplex virus -- Genetic aspects

DOI

10.15760/etd.6600

Physical Description

1 online resource (2, v, 83 pages)

Abstract

A regulatory element involved in the transcriptional activation of the major capsid protein (VP5) of herpes simplex virus type 1 was identified and characterized in this research project. Gel mobility shift assay with nuclear extracts from both uninfected and HSV-1 infected HeLa cells identified two major protein-DNA complexes involving the VP5 promoter. No viral specific complex found. DNase I and orthophenanthroline-Cu+ footprint analyses in the same laboratory revealed that the two complexes involve a single binding site, GGCCATCTTGAA, located between -64 and -75 bp relative to the VP5 cap site. To determine the function of this leaky-late binding site (LBS) in VP5 gene activation, mutated VP5 promoters with deletion and insertion around LBS were constructed and linked to a reporter gene, bacterial chloramphenicol acetyltransferase gene. The effect of mutations were tested in transient expression assay. Deletion of LBS resulted in seven to eight-fold reduction in the level of transactivation of the chloramphenicol acetyltransferase gene by superinfection with HSV-1 or by cotransfection of HSV immediate-early genes. These results indicated LBS is involved in the maximal transactivation of the VP5 gene. A search of published gene sequences found the homologs of LBS exist in a number of HSV-1 βγ promoters, and other viral promoters, as well as cellar promoters. Some of these homologs have found involved in the transcription regulation.

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Comments

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Persistent Identifier

https://archives.pdx.edu/ds/psu/27764

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