נגישות
menu      
Advanced Search
Syntax
Search...
Volcani treasures
About
Terms of use
Manage
Community:
אסיף מאגר המחקר החקלאי
Powered by ClearMash Solutions Ltd -
Reduced expression of the immediate-early protein IE0 enables efficient replication of Autographa californica multiple nucleopolyhedrovirus in poorly permissive Spodoptera littoralis cells
Year:
2003
Source of publication :
Journal of Virology
Authors :
Chejanovsky, Nor
;
.
Volume :
77
Co-Authors:
Lu, L., Entomology Department, Institute of Plant Protection, Volcani Center, POB 6, Bet Dagan 50250, Israel
Du, Q., Center for Cell Signaling, School of Medicine, University of Virginia, Charlottesville, VA 22908, United States
Chejanovsky, N., Entomology Department, Institute of Plant Protection, Volcani Center, POB 6, Bet Dagan 50250, Israel
Facilitators :
From page:
535
To page:
545
(
Total pages:
11
)
Abstract:
Infection of Spodoptera littoralis SL2 cells with the baculovirus Autographa californica multiple nucleopoly. hedrovirus (AcMNPV) results in apoptosis and low yields of viral progeny, in contrast to infection with S. littoralis nucleopolyhedrovirus (SINPV). By cotransfecting SL2 cells with AcMNPV genomic DNA and a cosmid library representing the complete SINPV genome, we were able to rescue AcMNPV replication and to isolate recombinant virus vAcSL2, which replicated efficiently in SL2 cells. Moreover, vAcSL2 showed enhanced infectivity for S. littoralis larvae compared to AcMNPV. The genome of vAcSL2 carried a 519-bp insert fragment that increased the distance between the TATA element and the transcriptional initiation site (CAGT) of immediate-early gene ie0. This finding correlated with low steady-state levels of IE0 and higher steady-state levels of IE1 (the product of the ie1 gene, a major AcMNPV transactivator, and a multifunctional protein) than of IE0. Mutagenesis of the ie0 promoter locus by insertion of the chloramphenical acetyltransferase (cat) gene yielded a new recombinant AcMNPV with replication properties identical to those of vAcSL2. Thus, the analysis indicated that increasing the steady-state levels of IE1 relative to IE0 should enable AcMNPV replication in SL2 cells. This suggestion was confirmed by constructing a recombinant AcMNPV bearing an extra copy of the ie1 gene under the control of the Drosophila hsp70 promoter. These results suggest that IE0 plays a role in the regulation of AcMNPV infection and show, for the first time, that significant improvement in the ability of AcMNPV to replicate in a poorly permissive cell line and organism can be achieved by Increasing the expression of the main multiple functional protein, IE1.
Note:
Related Files :
Animals
apoptosis
Autographa californica
Base Sequence
dimerization
Molecular Sequence Data
protein IE0
unclassified drug
virus replication
Show More
Related Content
More details
DOI :
10.1128/JVI.77.1.535-545.2003
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
29305
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:45
You may also be interested in
Scientific Publication
Reduced expression of the immediate-early protein IE0 enables efficient replication of Autographa californica multiple nucleopolyhedrovirus in poorly permissive Spodoptera littoralis cells
77
Lu, L., Entomology Department, Institute of Plant Protection, Volcani Center, POB 6, Bet Dagan 50250, Israel
Du, Q., Center for Cell Signaling, School of Medicine, University of Virginia, Charlottesville, VA 22908, United States
Chejanovsky, N., Entomology Department, Institute of Plant Protection, Volcani Center, POB 6, Bet Dagan 50250, Israel
Reduced expression of the immediate-early protein IE0 enables efficient replication of Autographa californica multiple nucleopolyhedrovirus in poorly permissive Spodoptera littoralis cells
Infection of Spodoptera littoralis SL2 cells with the baculovirus Autographa californica multiple nucleopoly. hedrovirus (AcMNPV) results in apoptosis and low yields of viral progeny, in contrast to infection with S. littoralis nucleopolyhedrovirus (SINPV). By cotransfecting SL2 cells with AcMNPV genomic DNA and a cosmid library representing the complete SINPV genome, we were able to rescue AcMNPV replication and to isolate recombinant virus vAcSL2, which replicated efficiently in SL2 cells. Moreover, vAcSL2 showed enhanced infectivity for S. littoralis larvae compared to AcMNPV. The genome of vAcSL2 carried a 519-bp insert fragment that increased the distance between the TATA element and the transcriptional initiation site (CAGT) of immediate-early gene ie0. This finding correlated with low steady-state levels of IE0 and higher steady-state levels of IE1 (the product of the ie1 gene, a major AcMNPV transactivator, and a multifunctional protein) than of IE0. Mutagenesis of the ie0 promoter locus by insertion of the chloramphenical acetyltransferase (cat) gene yielded a new recombinant AcMNPV with replication properties identical to those of vAcSL2. Thus, the analysis indicated that increasing the steady-state levels of IE1 relative to IE0 should enable AcMNPV replication in SL2 cells. This suggestion was confirmed by constructing a recombinant AcMNPV bearing an extra copy of the ie1 gene under the control of the Drosophila hsp70 promoter. These results suggest that IE0 plays a role in the regulation of AcMNPV infection and show, for the first time, that significant improvement in the ability of AcMNPV to replicate in a poorly permissive cell line and organism can be achieved by Increasing the expression of the main multiple functional protein, IE1.
Scientific Publication
You may also be interested in