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Here we attempt to reconstruct the sequence of events that led to the formation of three regulatory piRNA clusters, namely, 20A, 38C and flamenco in the Drosophila melanogaster genome. Both the 38C and flamenco clusters include inverted sequences, which potentially form double-stranded RNA hairpins. We present evidence in favor of the well-known hypothesis of piRNA clusters as “transposon traps”. According to this model, the presence of the only copy of the transposon in the genome indicates that its expression is suppressed by an RNA-interference mechanism immediately after the mobile element enters the piRNA cluster. We also discuss high the structural variability of piRNAs in Drosophila clusters and cases of horizontal transmobile elements between related species.

— The paper discusses the techniques which are currently implemented for vaccine production based on virus-like particles (VLPs). The factors which determine the characteristics of VLP monomers assembly are provided in detail. Analysis of the literature demonstrates that the development of the techniques of VLP production and immobilization of target antigens on their surface have led to the development of universal platforms which make it possible for virtually any known antigen to be exposed on the particle surface in a highly concentrated form. As a result, the focus of attention has shifted from the approaches to VLP production to the development of a precise interface between the organism’s immune system and the peptides inducing a strong immune response to pathogens or the organism’s own pathological cells. Immunome-specified methods for vaccine design and the prospects of immunoprophylaxis are discussed. Certain examples of vaccines against viral diseases and cancers are considered.

The structural protein (Gag) of Drosophila retrovirus gypsy contains capsid and nucleocapsid domains. Gag forms virus-like particles in a bacterial cell; furthermore, its capsid alone is able to form aggregates. However, aggregates assembled from the capsid vary in size and are less organized than particles formed by a full-length Gag. The nucleocapsid determines the organization and structure of the particles, which is ensured by the amino acid residues at its N-terminal (a nucleocapsid proximal part). The assembly of the particle occurs in the presence of any RNAs or single-stranded DNA oligonucleotides.

Amino acid sequence of the drosophila retrovirus MDG4 (gypsy) structural protein Gag does not contain a canonical motif known for the majority of vertebrate retroviruses. Moreover, protein translation can theoretically begin with two separated initiation codons located within its unique open reading frame. We designed constructs for expression of two theoretically possible variants of Gag polypeptide and investigated the ability of each product to form virus-like particles in the bacterial cell, i.e., in the absence of eukaryotic cell factors. The results obtained showed that both variants of the gypsy protein Gag form globular particles in the bacterial cell.

Since retrovirus-like particles of gypsy (mdg4) are capable of interspecific transfer, other Drosophila melanogaster gypsy-related retrotransposons were tested for this property. As a donor and a recipient, D. melanogaster and D. virilis cultured cells were used. Recipient cell DNA was analyzed with probes directed to mdg1, mdg3, 17.6, 297, 412, or B104/roo. Transfer was demonstrated for mdg3, which lacks env. The possible mechanism of transfer is discussed.

To study the mechanism of precise excision ofgypsy from genomic sites, the integrase domain ofgypsy pol was cloned and expressed inEscherichia coli. The endonuclease activity of recombinant integrase was assayed with synthetic substrates corresponding to 3′-U5 ofgypsy LTR and to the known genomic insertion sites ofgypsy. Integrase nicked the 5′-A ⇓ YR-3′ triplet in the (+) strand of the double-stranded substrates; cleavage of a single-stranded substrate was nonspecific. Cleavage proved to be affected by the local conformation of the substrate: the (+) strand was cleaved more efficiently when the (−) strand had an unpaired base in the triplet and was not cleaved when the (−) strand was interrupted or branched. The triplet corresponded to the consensus region ofgypsy insertion (5′-YRYR ⇓ YR-3′), the site of cleavagein vitro coinciding with the site of insertionin vivo. The unique mechanism ofgypsy excision was assumed to depend to a great extent on the enzymic properties of its integrase.