| Abstract: | The Autographa californica multinucleocapsid nuclear polyhedrosis virus (AcMNPV) ie-1 gene product (IE-1) is thought to play a central role in stimulating early viral transcription. IE-1 has been demonstrated to activate several early viral gene promoters and to negatively regulate the promoters of two other AcMNPV regulatory genes, ie-0 and ie-2. Our results indicate that IE-1 negatively regulates the expression of certain genes by binding directly, or as part of a complex, to promoter regions containing a specific IE-1-binding motif (5‘-ACBYGTAA-3‘) near their mRNA start sites. The IE-1 binding motif was also found within the palindromic sequences of AcMNPV homologous repeat (hr) regions that have been shown to bind IE-1. The role of this IE-1 binding motif in the regulation of the ie-2 and pe-38 promoters was examined by introducing mutations in these promoters in which the central 6 bp were replaced with Bg/II sites. GUS reporter constructs containing ie-2 and pe-38 promoter fragments with and without these specific mutations were cotransfected into Sf9 cells with various amounts of an ie-1-containing plasmid (ple-1). Comparisons of GUS expression produced by the mutant and wild-type constructs demonstrated that the IE-1 binding motif mediated a significant decrease in expression from the ie-2 and pe-38 promoters in response to increasing pIe-1 concentrations. Electrophoretic mobility shift assays with pIe-1-transfected cell extracts and supershift assays with IE-1- specific antiserum demonstrated that IE-1 binds to promoter fragments containing the IE-1 binding motif but does not bind to promoter fragments lacking this motif. |
| Genre: | Article |
| ProdID: | 70019722 |
| Citation Author: | Leisy, D. J.; Rasmussen, C.; Owusu, E. O.; Rohrmann, G. F. |
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| Citation End Page: | 5094 |
| Citation Issue: | 7 |
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| Citation Language: | English |
| Citation Larger Work Title: | Journal of Virology |
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| Citation Number Of Pages: | 7 |
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| Citation Search Results Text: | A mechanism for negative gene regulation in Autographa californica multinucleocapsid nuclear polyhedrosis virus; 1997; Article; Journal; Journal of Virology; Leisy, D. J.; Rasmussen, C.; Owusu, E. O.; Rohrmann, G. F. |
| Citation Start Page: | 5088 |
| Citation Volume: | 71 |
| Citation Year: | 1997 |
| Type: | citation/reference |
| Text: | A mechanism for negative gene regulation in Autographa californica multinucleocapsid nuclear polyhedrosis virus; 1997; Article; Journal; Journal of Virology; Leisy, D. J.; Rasmussen, C.; Owusu, E. O.; Rohrmann, G. F. |
| URL (THUMBNAIL): | http://pubs.er.usgs.gov/thumbnails/outside_thumb.jpg |
| Date Other: | Wed, 1 Jan 1997 00:00 -0600 |
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