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Theie2 gene ofAutographa californica nuclear polyhedrosis virus (AcM NPV) is known to transactivate transient expression from viral promoters in a host cell-specific manner. We report that transfection ofSpodoptera frugiperda (SF-21) cells withie2 was sufficient to arrest the cell cycle, resulting in the accumulation of enlarged cells with abnormally high DNA contents. By 72 h posttransfection, more than 50% ofie2 -transfected cells had DNA contents greater than 4N. There was no evidence of mitotic spindle formation in these cells, and expression ofie2 appeared to block cell cycle progression in S phase. Severalie2 mutants were analyzed to further define the region of IE2 responsible for arresting the cell cycle. Analysis of these mutants showed that deletion of the RING finger motif eliminated the ability of IE2 to arrest the cell cycle but did not affect its ability to transactivate theie1 promoter. Moreover, mutation of a single conserved cysteine (C251) of the RING finger motif abolished the ability of IE2 to block cell cycle progression but had no apparent effect on itstrans -regulatory activity. In contrast, a mutant of IE2 containing a deletion of residues 94 to 173 was able to block cell division but lackedtrans -regulatory activity. Thus, the ability of IE2 to arrest the cell cycle depended on the integrity of the RING finger motif and was distinct from and independent of its ability totrans -activate theie1 promoter. IE2 also arrested the division of cells derived from other insect species,Trichoplusia ni (TN-368 and BTI-TN-5B1-4) andHelicoverpa zea (Hz-AM1).

Role of Baculovirus IE2 and Its RING Finger in Cell Cycle Arrest