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

<jats:title>ABSTRACT</jats:title> <jats:p> The genome of the baculovirus <jats:italic>Autographa californica</jats:italic> nuclear polyhedrosis virus (Ac <jats:italic>M</jats:italic> NPV) contains two homologues, <jats:italic>orf145</jats:italic> and <jats:italic>orf150</jats:italic> , of the <jats:italic>Heliothis armigera</jats:italic> 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 <jats:italic>orf145</jats:italic> and <jats:italic>orf150</jats:italic> were deleted from the Ac <jats:italic>M</jats:italic> NPV genome, singly or together, and these deletion mutants were assessed for oral infectivity both in <jats:italic>Trichoplusia ni</jats:italic> and <jats:italic>Heliothis virescens</jats:italic> larvae. Deletion of Ac145 led to a small but significant drop in infectivity (sixfold) compared to wild-type (wt) Ac <jats:italic>M</jats:italic> NPV for <jats:italic>T. ni</jats:italic> but not for <jats:italic>H. virescens</jats:italic> . 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 <jats:italic>H. virescens</jats:italic> . Intrahemocoelic injection of budded virus from the double-deletion virus into <jats:italic>H. virescens</jats:italic> larvae is as infectious to this host as wt budded virus, indicating that Ac145 and Ac150 play a role in primary oral infection of Ac <jats:italic>M</jats:italic> NPV, the extent of which is host dependent. </jats:p>