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Journal of Virology, June 2004, p. 6439-6448, Vol. 78, No. 12
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.12.6439-6448.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of Two Autographa californica Nucleopolyhedrovirus Proteins, Ac145 and Ac150, Which Affect Oral Infectivity in a Host-Dependent Manner

Renee Lapointe,1,2 Holly J. R. Popham,3 Ursula Straschil,1 David Goulding,1 David R. O'Reilly,1,4 and Julie A. Olszewski1*

Department of Biological Sciences, Imperial College, London SW7 2AZ,1 Syngenta, Jealotts Hill International Research Centre, Bracknell, Berks RG42 6EY, United Kingdom,4 Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, Sainte-Foy, Québec, Canada G1V 4C7,2 Biological Control of Insects Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Columbia, Missouri 652033

Received 4 November 2003/ Accepted 3 February 2004

The genome of the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) contains two homologues, orf145 and orf150, of the Heliothis armigera Entomopoxvirus (HaEPV) 11,000-kDa gene. Polyclonal antibodies raised against the Ac145 or Ac150 protein were utilized to demonstrate that they are expressed from late to very late times of infection and are within the nuclei of infected Sf-21 cells. Transmission electron microscopy coupled with immunogold labeling of Ac145 found this protein within the nucleus in areas of nucleocapsid assembly and maturation, along with some association with the enveloped bundles of virions within the developing occlusion bodies (OBs). Ac150 was found to be mainly associated with enveloped bundles of virions within OBs and also with those not yet occluded. Both Ac145 and Ac150 were found to be present in budded virus as well as OBs. Both orf145 and orf150 were deleted from the AcMNPV genome, singly or together, and these deletion mutants were assessed for oral infectivity both in Trichoplusia ni and Heliothis virescens larvae. Deletion of Ac145 led to a small but significant drop in infectivity (sixfold) compared to wild-type (wt) AcMNPV for T. ni but not for H. virescens. Deletion of Ac150 alone had no effect on infectivity of the virus for either host. However, deletion of both Ac145 and Ac150 gave a recombinant virus with a drastic (39-fold) reduction in infectivity compared to wt virus for H. virescens. Intrahemocoelic injection of budded virus from the double-deletion virus into H. virescens larvae is as infectious to this host as wt budded virus, indicating that Ac145 and Ac150 play a role in primary oral infection of AcMNPV, the extent of which is host dependent.

* Corresponding author. Mailing address: Department of Biological Sciences, SAFB, South Kensington Campus, London SW7 2AZ, United Kingdom. Phone: 44 0 20 7594 5376. Fax: 44 0 20 7584 2056. E-mail: j.olszewski{at}imperial.ac.uk.

Journal of Virology, June 2004, p. 6439-6448, Vol. 78, No. 12
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.12.6439-6448.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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