Expression of PE38 and IE2, viral members of the C3HC4 finger family, during baculovirus infection: PE38 and IE2 localize to distinct nuclear regions.
The pe38 gene of Autographa californica nuclear polyhedrosis virus represents one of the major early transcripts after viral infection. The function of the pe38 protein, which contains a C3HC4 zinc finger motif, is still not understood. We have raised polyclonal antiserum against the pe38 protein, PE38, produced in bacteria to investigate pe38 expression in the course of infection. A approximately 38-kDa polypeptide is first detectable at 2 h postinfection and decreases rapidly after 24 h. During the late phases of infection, a smaller protein of approximately 20 kDa which cross-reacts with the PE38-specific antiserum is visible at a constant level until 120 h postinfection. Since the pe38 gene shares a divergent promoter unit with the ie2 gene (formerly IEN), we have compared the expressions of the two genes. Polyclonal antibodies were raised against the bacterially expressed ie2 protein. The temporal expression pattern of the approximately 49-kDa ie2 protein is comparable to that of the approximately 38-kDa pe38 protein. Furthermore, both proteins are present in the nuclear fraction of A. californica nuclear polyhedrosis virus-infected Spodoptera frugiperda cells, but the approximately 38-kDa pe38 protein is also detectable in the cytoplasm while the smaller protein of approximately 20 kDa is exclusively present in the cytoplasmic fraction. Immunofluorescence analysis reveals that PE38 and IE2 localize to distinct regions within the nucleus mainly detected after transfection of pe38- and ie2-expressing constructs.
Krappa
R
R
Institute of Genetics, University of Cologne, Germany.
Roncarati
R
R
Knebel-Mörsdorf
D
D
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers; D004279:DNA, Viral / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D018411:Spodoptera; D015534:Trans-Activators / Q000096:biosynthesis* / Q000302:isolation & purification; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins; D014779:Virus Replication; D016335:Zinc Fingers
C095109:Ac-PE38 protein, Autographa californica nucleopolyhedrovirus; D017931:DNA Primers; D004279:DNA, Viral; D017874:Immediate-Early Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C072773:IE2 protein, polyhedrosis virus; D016590:Aphidicolin; D003513:Cycloheximide