Abstract
At present, the details of lamina alterations after baculovirus infection remain elusive. In this study, a lamin gene in the Sf9 cell line of Spodoptera frugiperda was cloned. The open reading frame (orf) of the Sf9 lamin was 1860 bp and encoded a protein with a molecular weight of 70 kDa. A transfection assay with a red fluorescence protein (rfp)-lamin fusion protein indicated that Sf9 lamin was localized in the nuclear rim. Transmission electron microscopy observations indicated that Autographa californica multiple nucleopolyhedrovirus (AcMNPV) nucleocapsids may pass through the nuclear envelope. Immunofluorescence assay indicated that the lamina showed a ruffled staining pattern with the formation of invaginations in the Sf9 cells infected with AcMNPV, while it was evenly distributed at the nuclear periphery of mock-infected cells. Western blotting results indicated that the total amount of lamin in the baculovirus-infected Sf9 cells was significantly decreased compared with the mock-infected cells. These results imply that AcMNPV infection induces structural and biochemical rearrangements of lamina of Sf9 cells.
Highlights
The nuclear membrane consists of the outer and inner nuclear membranes separated by the perinuclear space, the nuclear pore complexes, and the nuclear lamina
A 70 kDa protein band was observed from the total protein lysates of Sf9 cells (Figure 1A)
The lamin of Sf9 cells is indistinguishable in its molecular weight from Drosophila lamin Dm0, suggesting that these Sf9 cells do express the lamin
Summary
The nuclear membrane consists of the outer and inner nuclear membranes separated by the perinuclear space, the nuclear pore complexes, and the nuclear lamina. (HHV-1) can disrupt the lamina to promote nucleocapsid egress from the nucleus [8]. HHV-1 recruits cellular protein kinase C to phosphorylate emerin and lamin to induce the disruption of nuclear lamina [9]. Wheat germ agglutinin-gold labeling experiments demonstrated that nucleocapsids move from the prominent pore in the nuclear membrane to the cytoplasm [15]. These data provide evidence that baculoviruses may pass through the nuclear membrane and enter the cytoplasm. We cloned the orf sequence of lamin (similar to the Drosophila melanogaster nuclear lamin Dm0) in Sf9 cells and observed some of the changes in Sf9 lamin following baculovirus infection
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