Mutagenesis of baculoviruses by host mobile elements occurs spontaneously and frequently during propagation of the viruses in Lepidopteran cell cultures. Most of the transposons identified as insertions in baculovirus genomes are relatively small Class II elements that exhibit a remarkable specificity for TTAA target sites. We have developed a transposition assay to analyze the movement of these TTAA-specific Lepidopteran transposons using the baculovirus genome as a target and a lacZ gene under control of the polyhedrin gene promoter as a selective marker for the transposon. This assay provides the first demonstration that a Lepidopteran transposon is capable of transposing while carrying a marker gene in insect cells. The data generated from this assay provide strong evidence that IFP2 encodes a protein that facilitates its own movement. This element may be used in a manner analogous to the P-element to mobilize genes in at least some Lepidopteran insect cells. Transposon tagging within the baculovirus genome identified several known genes and two previously undescribed open reading frames as nonessential to in vitro replication of the virus.

BMS-378806 is a recently discovered small-molecule human immunodeficiency virus type 1 (HIV-1) attachment inhibitor with good antiviral activity and pharmacokinetic properties. Here, we demonstrate that the compound targets viral entry by inhibiting the binding of the HIV-1 envelope gp120 protein to cellular CD4 receptors via a specific and competitive mechanism. BMS-378806 binds directly to gp120 at a stoichiometry of approximately 1:1, with a binding affinity similar to that of soluble CD4. The potential BMS-378806 target site was localized to a specific region within the CD4 binding pocket of gp120 by using HIV-1 gp120 variants carrying either compound-selected resistant substitutions or gp120-CD4 contact site mutations. Mapping of resistance substitutions to the HIV-1 envelope, and the lack of compound activity against a CD4-independent viral infection confirm the gp120-CD4 interactions as the target in infected cells. BMS-378806 therefore serves as a prototype for this new class of antiretroviral agents and validates gp120 as a viable target for smallmolecule inhibitors.

We have analysed TFP3 transposable elements from five independently isolated FP mutants of the Autographa californica nuclear polyhderosis virus (AcMNPV). We also analysed genomic copies of TFP3 elements amplified from the DNAs of the Trichoplusia ni cell line (TN-368) and T. ni larvae using the polymerase chain reaction (PCR). The sequences of all the newly isolated TFP3 elements closely resemble the previously described TFP3/1 element. Each of the transposons isolated from the virus mutants duplicated a TTAA tetranucleotide target site upon insertion into the viral genome. Four of these TFP3 elements transposed into three different 'TTAA' target sites within the 25 K gene (FP locus, map units 36-37 of AcMNPV). The fifth TFP3 element inserted at a 'TTAA' site within the AcMNPV Hin dIII-E fragment. One genomic TFP3 element, amplified from the TN-368 cell line DNA by an inverse PCR method, duplicated a 'TTAA' tetranucleotide target site that is present only once in the homologous larval DNA sequence. These data suggest that mobilization of TFP3 into both viral and cellular sites is identical in specificity and mechanism.

No abstract