This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Glocker, B Articles by Rohrmann, G F Search for Related Content PubMed PubMed Citation Articles by Glocker, B Articles by Rohrmann, G F

In vitro transcription from baculovirus late gene promoters: accurate mRNA initiation by nuclear extracts prepared from infected Spodoptera frugiperda cells.

Department of Agricultural Chemistry, Oregon State University, Corvallis 97331-7301.

ABSTRACT

Extracts prepared from nuclei of Autographa californica nuclear polyhedrosis virus-infected Spodoptera frugiperda cells were shown to support in vitro transcription from baculovirus late gene promoters. In vitro transcription was optimized for the late promoter of the 39K gene. The Mg2+ concentration was critical; concentrations higher than 1 to 2 mM did not support late transcription. Additional conditions included template (40 micrograms/ml), extract (2.5 mg/ml), and incubation time (25 min). Using a combination of runoff assays and high-resolution primer extension analyses, this system was shown to accurately initiate transcription from a variety of baculovirus late gene promoters, including those from the 39K and p39/capsid late genes and the hyperexpressed p10 and polyhedrin very late genes. In vitro transcription from the 39K late promoter was resistant to high concentrations of both alpha-amanitin (100 micrograms/ml) and tagetitoxin (4,000 U/ml), suggesting that neither RNA polymerase II nor III is responsible for the transcription of baculovirus late genes.

This article has been cited by other articles:

• Knebel-Morsdorf, D., Quadt, I., Li, Y., Montier, L., Guarino, L. A. (2006). Expression of Baculovirus Late and Very Late Genes Depends on LEF-4, a Component of the Viral RNA Polymerase Whose Guanyltransferase Function Is Essential.. J. Virol. 80: 4168-4173 [Abstract] [Full Text]  
• Titterington, J. S., Nun, T. K., Passarelli, A. L. (2003). Functional dissection of the baculovirus late expression factor-8 gene: sequence requirements for late gene promoter activation. J. Gen. Virol. 84: 1817-1826 [Abstract] [Full Text]  
• Lin, G., Slack, J. M., Blissard, G. W. (2001). Expression and localization of LEF-11 in Autographa californica nucleopolyhedrovirus-infected Sf9 cells. J. Gen. Virol. 82: 2289-2294 [Abstract] [Full Text]  
• Evans, J., Rosenblatt, G., Leisy, D., Rohrmann, G. (1999). Characterization of the interaction between the baculovirus ssDNA- binding protein (LEF-3) and putative helicase (P143). J. Gen. Virol. 80: 493-500 [Abstract]  
• Jin, J., Dong, W., Guarino, L. A. (1998). The LEF-4 Subunit of Baculovirus RNA Polymerase Has RNA 5'-Triphosphatase and ATPase Activities. J. Virol. 72: 10011-10019 [Abstract] [Full Text]  
• Guarino, L. A., Xu, B., Jin, J., Dong, W. (1998). A Virus-Encoded RNA Polymerase Purified from Baculovirus-Infected Cells. J. Virol. 72: 7985-7991 [Abstract] [Full Text]  
• Mans, R. M. W., Knebel-Morsdorf, D. (1998). In Vitro Transcription of pe38/Polyhedrin Hybrid Promoters Reveals Sequences Essential for Recognition by the Baculovirus-Induced RNA Polymerase and for the Strength of Very Late Viral Promoters. J. Virol. 72: 2991-2998 [Abstract] [Full Text]