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.

Rights

In Copyright. URI: http://rightsstatements.org/vocab/InC/1.0/ This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).

Comments

If you are the rightful copyright holder of this dissertation or thesis and wish to have it removed from the Open Access Collection, please submit a request to pdxscholar@pdx.edu and include clear identification of the work, preferably with URL.

Persistent Identifier

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

Included in

Biology Commons

Share

COinS