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Journal of Virology, August 2005, p. 10578-10588, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10578-10588.2005

The Lymantria dispar Nucleopolyhedrovirus Enhancins Are Components of Occlusion-Derived Virus

James M. Slavicek* and Holly J. R. Popham{dagger}

USDA Forest Service, Northeastern Research Station, Forestry Sciences Laboratory, 359 Main Road, Delaware, Ohio 43015

Received 15 February 2005/ Accepted 13 May 2005

Enhancins are metalloproteinases, first identified in granuloviruses, that can enhance nucleopolyhedrovirus (NPV) potency. We had previously identified two enhancin genes (E1 and E2) in the Lymantria dispar multinucleocapsid NPV (LdMNPV) and showed that both were functional. For this study, we have extended our analysis of LdMNPV enhancin genes through an immunocytochemical analysis of E1 and E2 expression and localization. E1 and E2 peptide antibodies recognized proteins of ~84 kDa and 90 kDa, respectively, on Western blots of extracts from L. dispar 652Y cells infected with wild-type virus. The 84- and 90-kDa proteins were first detected at 48 h postinfection (p.i.) and were present through 96 h p.i. E1 and E2 peptide antibodies detected E1 and E2 in polyhedron extracts, and the antibodies were shown to be specific for E1 and E2, respectively, through the use of recombinant virus strains lacking the enhancin genes. E1 and E2 were further localized to occlusion-derived virus (ODV). The enhancins were not found in budded virus. Immunoelectron microscopy indicated that E1 and E2 were present in ODV envelopes and possibly in nucleocapsids. Fractionation studies with several detergents suggested that the enhancins were present in ODV envelopes in association with nucleocapsids. In contrast, enhancins in granuloviruses are located within the granulin matrix. The presence of LdMNPV enhancins within ODV provides a position for the proteins to interact directly on the peritrophic membrane as ODV traverses this host defense barrier.

* Corresponding author. Mailing address: USDA Forest Service, 359 Main Road, Delaware, OH 43015. Phone: (740) 368-0033. Fax: (740) 368-0152. E-mail: jslavicek{at}fs.fed.us.

{dagger} Present address: USDA Agricultural Research Service, Biological Control of Insects Research Laboratory, 1503 S. Providence Rd., Columbia, MO 65203.

Journal of Virology, August 2005, p. 10578-10588, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10578-10588.2005




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