TY - JOUR
T1 - Expression of the insect parvovirus GmDNV in vivo
T2 - The structural and nonstructural proteins are encoded by opposite DNA strands
AU - Gross, Orit
AU - Tal, Jacov
N1 - Funding Information:
We are grateful to Peter Tijssen for an initial gift of GmDNV and for the pGmDNV clone, Michal Mincberg for expert technical assistance throughout the course of this work, and Joe Corsini for critical reading of the manuscript. This study was supported by grants from the National Council for Research and Development, Israel, and GSF, München, Germany; and from the US Agency for Research and Development (AID-CDR).
PY - 2000/1/1
Y1 - 2000/1/1
N2 - The nucleotide sequence of GmDNV, an insect parvovirus, reveals large open reading frames (ORFs) on both strands of the viral replicative form DNA. Previously, we identified two viral transcripts within the polyadenylated RNA fraction of infected host larvae (Gross et al., 1990, J. Invertebr. Pathol. 56, 175-180). In this work we used hybridization of single-stranded, unidirectional probes to RNA blots to show that the two transcripts, synthesized in vivo in GmDNV-infected Galleria mellonella larvae, are of antiparallel orientation. To determine their coding specificities, polyadenylated RNAs were isolated from hybrids with DNA from the left and right halves of the viral genome and translated in a rabbit reticulocyte system. The 'right,' 2.4-kb hybrid-selected RNA was shown to direct the synthesis of four polypeptides that comigrated with the four viral capsid proteins and were immunoprecipitated with anti-GmDNV serum. Translation of the 'left,' 1.8-kb RNA yielded three polypeptides, none of which was detected among the vital capsid proteins. This type of expression strategy is unique among vertebrate and most invertebrate parvoviruses, which use only one DNA strand to encode all their proteins. On the other hand, the basic organization of parvoviruses, in which the regulatory and structural proteins are encoded, respectively, by two clusters of ORFs located at the left and right halves of the genome, is conserved. (C) 2000 Academic Press.
AB - The nucleotide sequence of GmDNV, an insect parvovirus, reveals large open reading frames (ORFs) on both strands of the viral replicative form DNA. Previously, we identified two viral transcripts within the polyadenylated RNA fraction of infected host larvae (Gross et al., 1990, J. Invertebr. Pathol. 56, 175-180). In this work we used hybridization of single-stranded, unidirectional probes to RNA blots to show that the two transcripts, synthesized in vivo in GmDNV-infected Galleria mellonella larvae, are of antiparallel orientation. To determine their coding specificities, polyadenylated RNAs were isolated from hybrids with DNA from the left and right halves of the viral genome and translated in a rabbit reticulocyte system. The 'right,' 2.4-kb hybrid-selected RNA was shown to direct the synthesis of four polypeptides that comigrated with the four viral capsid proteins and were immunoprecipitated with anti-GmDNV serum. Translation of the 'left,' 1.8-kb RNA yielded three polypeptides, none of which was detected among the vital capsid proteins. This type of expression strategy is unique among vertebrate and most invertebrate parvoviruses, which use only one DNA strand to encode all their proteins. On the other hand, the basic organization of parvoviruses, in which the regulatory and structural proteins are encoded, respectively, by two clusters of ORFs located at the left and right halves of the genome, is conserved. (C) 2000 Academic Press.
KW - Densovirus
KW - Gene expression
KW - In vitro translation
UR - http://www.scopus.com/inward/record.url?scp=0034072051&partnerID=8YFLogxK
U2 - 10.1006/jipa.1999.4904
DO - 10.1006/jipa.1999.4904
M3 - Article
C2 - 10772325
AN - SCOPUS:0034072051
SN - 0022-2011
VL - 75
SP - 126
EP - 132
JO - Journal of Invertebrate Pathology
JF - Journal of Invertebrate Pathology
IS - 2
ER -