An element downstream of the transcription start site is required for activation of Bombyx mori nucleopolyhedrovirus bro-c promoter

An element downstream of the transcription start site is required for activation of Bombyx mori... Bombyx mori nucleopolyhedrovirus (BmNPV) contains five baculovirus repeated ORF ( bro ) genes. We have previously reported that all of these genes ( bro-a, b, c, d and e ) are transcribed as early genes and require viral factor(s) for their expression. In this study, we investigated the mechanism of promoter activation of the bro-c gene. Transient expression assays using genomic libraries of BmNPV indicated that the baculoviral trans -regulator IE-1 is responsible for activating the bro-c promoter. To identify essential site(s) for promoter activation, mutations were introduced to the promoter region of bro-c . Interestingly, it was shown that the pentanucleotide sequence CACGC located 30 nucleotides downstream of the RNA start site was essential for bro-c promoter activation. In addition, the RNA start site and the spacing between the RNA start site and CACGC were also required for promoter activation. By introducing a CACGC sequence into the corresponding region of the bro-b promoter, which is not normally trans-activated by IE-1, we demonstrated that this pentanucleotide motif has the ability to confer trans -activation by IE-1 on a promoter. Gel retardation experiments also showed a sequence-specific DNA binding protein induced by baculovirus infection interacts with the CACGC motif. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

An element downstream of the transcription start site is required for activation of Bombyx mori nucleopolyhedrovirus bro-c promoter

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Publisher
Springer Journals
Copyright
Copyright © 2001 by Springer-Verlag/Wien
Subject
Legacy
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s007050170158
Publisher site
See Article on Publisher Site

Abstract

Bombyx mori nucleopolyhedrovirus (BmNPV) contains five baculovirus repeated ORF ( bro ) genes. We have previously reported that all of these genes ( bro-a, b, c, d and e ) are transcribed as early genes and require viral factor(s) for their expression. In this study, we investigated the mechanism of promoter activation of the bro-c gene. Transient expression assays using genomic libraries of BmNPV indicated that the baculoviral trans -regulator IE-1 is responsible for activating the bro-c promoter. To identify essential site(s) for promoter activation, mutations were introduced to the promoter region of bro-c . Interestingly, it was shown that the pentanucleotide sequence CACGC located 30 nucleotides downstream of the RNA start site was essential for bro-c promoter activation. In addition, the RNA start site and the spacing between the RNA start site and CACGC were also required for promoter activation. By introducing a CACGC sequence into the corresponding region of the bro-b promoter, which is not normally trans-activated by IE-1, we demonstrated that this pentanucleotide motif has the ability to confer trans -activation by IE-1 on a promoter. Gel retardation experiments also showed a sequence-specific DNA binding protein induced by baculovirus infection interacts with the CACGC motif.

Journal

Archives of VirologySpringer Journals

Published: Mar 1, 2001

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