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Title: | The Function of DNA Binding in the Regulation of HSV-1 Gene Expression by ICP4 |
Other Titles: | Regulation of HSV-1 Gene Expression by ICP4 |
Authors: | Koop, Karen |
Advisor: | Smiley, James |
Department: | Biology |
Keywords: | DNA;HSV-1;gene;expression |
Publication Date: | Apr-1994 |
Abstract: | Herpes simplex virus is an important model in the study of temporally regulated gene expression in eukaryotic cells. Three classes of genes - immediate early, early, and late - are sequentially expressed during the course of lytic infection. One immediate early gene product, ICP4, is required for transactivation of most early and late genes; it is also implicated in repression of immediate early gene expression. ICP4's mechanism(s) of action is/are not yet understood; although ICP4 binds to specific sequences of DNA, whether this is necessary for transregulation by ICP4 is not clear. To gain a better understanding of how the ability of ICP4 to bind DNA relates to its transregulatory activities, I introduced ICP4 binding sites into a simple model promoter within the viral genome. Two sets of construct were made in which an ICP4 binding site (or mutant site) was placed either downstream or upstream of a TATA box, reproducing the spacing found in (i) the native 𝘐𝘊𝘗4 promoter and (ii) the native 𝘐𝘊𝘗0 promoter (respectively). The promoter of HSV-1 𝘜𝘓24𝘣 (a nonessential gene in tissue culture) was replaced with these model promoters and levels of transcripts accumulating from these constructs during lytic infection assayed by primer extension. I found that an ICP4 site placed either upstream or downstream of a TATA box shifted kinetics of expression from E/leaky L to true L. Neither the strength of the TATA box nor the helical orientation of the ICP4 binding site with respect to the TATA box affected this result. |
URI: | http://hdl.handle.net/11375/22979 |
Appears in Collections: | Digitized Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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koop_karen_1994Apr_masters.pdf | 14.84 MB | Adobe PDF | View/Open |
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