Abstract
Autographa californica nuclear polyhedrosis virus (AcMNPV) is a double-stranded DNA virus that expresses several immediate-early genes under the control of different promoters. The expression of one of these transcription units, IE-N, is shown here, by a transient expression assay, to be regulated by both cis- and trans-acting viral elements. The steady-state levels of IE-N mRNA were very abundant soon after infection but were nearly undetectable during the late phase of the viral life cycle. Analysis of the transient expression of a reporter construct driven by the IE-N promoter (IE-NCAT) was conducted to define viral elements which regulate IE-N gene expression. Viral enhancer hr1 and two immediate-early genes, IE-1 and IE-N, were shown to affect relative levels of reporter enzyme activity produced by IE-NCAT. The hr1 enhancer stimulated the expression of IE-NCAT, independent of orientation and position relative to the promoter and in the absence of any trans-acting viral factors. Regulation of IE-NCAT expression by the IE-1 and IE-N genes required less than 290 bp of promoter sequences upstream of the site of transcription initiation and was not dependent upon the hr1 enhancer. Coexpression of the IE-N gene had an autostimulatory effect upon IE-NCAT activity, whereas coexpression of the IE-1 gene reduced levels of reporter activity. The levels of reporter activity measured upon coexpression of either immediate-early gene with IE-NCAT linked to the hr1 enhancer appear to be the combined result of both cis- and trans-regulatory elements influencing expression from IE-NCAT. These results suggest that IE-N gene expression in baculovirus infection may be influenced by the concerted activity of three AcMNPV regulatory elements.
Full text
PDF![945](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/c0c3864c2496/jvirol00045-0399.png)
![946](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/6fe6ca37a224/jvirol00045-0400.png)
![947](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/469e1d22883b/jvirol00045-0401.png)
![948](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/a200bd2f73fd/jvirol00045-0402.png)
![949](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/11d31754a3e4/jvirol00045-0403.png)
![950](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/2e5bba85b552/jvirol00045-0404.png)
![951](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ec1/239836/35034d7ad08c/jvirol00045-0405.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
- Carson D. D., Guarino L. A., Summers M. D. Functional mapping of an AcNPV immediately early gene which augments expression of the IE-1 trans-activated 39K gene. Virology. 1988 Feb;162(2):444–451. doi: 10.1016/0042-6822(88)90485-0. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Chisholm G. E., Henner D. J. Multiple early transcripts and splicing of the Autographa californica nuclear polyhedrosis virus IE-1 gene. J Virol. 1988 Sep;62(9):3193–3200. doi: 10.1128/jvi.62.9.3193-3200.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cochran M. A., Faulkner P. Location of Homologous DNA Sequences Interspersed at Five Regions in the Baculovirus AcMNPV Genome. J Virol. 1983 Mar;45(3):961–970. doi: 10.1128/jvi.45.3.961-970.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
- Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guarino L. A., Gonzalez M. A., Summers M. D. Complete Sequence and Enhancer Function of the Homologous DNA Regions of Autographa californica Nuclear Polyhedrosis Virus. J Virol. 1986 Oct;60(1):224–229. doi: 10.1128/jvi.60.1.224-229.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guarino L. A., Summers M. D. Functional mapping of Autographa california nuclear polyhedrosis virus genes required for late gene expression. J Virol. 1988 Feb;62(2):463–471. doi: 10.1128/jvi.62.2.463-471.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guarino L. A., Summers M. D. Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. J Virol. 1986 Feb;57(2):563–571. doi: 10.1128/jvi.57.2.563-571.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guarino L. A., Summers M. D. Interspersed Homologous DNA of Autographa californica Nuclear Polyhedrosis Virus Enhances Delayed-Early Gene Expression. J Virol. 1986 Oct;60(1):215–223. doi: 10.1128/jvi.60.1.215-223.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guarino L. A., Summers M. D. Nucleotide sequence and temporal expression of a baculovirus regulatory gene. J Virol. 1987 Jul;61(7):2091–2099. doi: 10.1128/jvi.61.7.2091-2099.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nissen M. S., Friesen P. D. Molecular analysis of the transcriptional regulatory region of an early baculovirus gene. J Virol. 1989 Feb;63(2):493–503. doi: 10.1128/jvi.63.2.493-503.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]