[Structure and Evolution of DNA Transposons of the L31 Superfamily in Bivalves].

The mobile genetic elements IS630/Tc 1/mariner (ITm) are widespread DNA transposons that make a significant contribution to the evolution of eukaryotic genomes. With the start of large-scale application of next-generation sequencing (NGS) technologies and the emergence of many new whole genome sequences of organisms in nucleotide sequence collections, the ITm elements have been identified in most taxa of the eukaryotic tree of life. Although ITm diversity has been studied in detail, new elements are still found, thus expanding the respective DNA transposon group and calling for review of its classification. Bivalve L31 elements were for the first time analyzed in detail to describe their structures, diversity, distribution, and phylogenetic position among the ITm elements. The L31 transposons were found to form an independent superfamily of an ancient origin within the ITm group. Rather high diversity was observed within the L31 clade; i.e., five phylogenetic clusters were identified. In mollusks, the L31 transposons have been detected only in the subclass Autobranchia and predominate in diversity and number in the infraclass Pteriomorphia. A protein encoded by open reading frame 2 (ORF2) was shown to be an integral structural component of almost all full-length L31 elements. The results provide for a better understanding of the evolution of particular ITm transposons. Further study of the L31 transposons in other taxa (cnidarians) and functional investigation of the ORF2 protein product will help to better understand the evolution of DNa transposons, the mechanisms of their horizontal transfer, and their contribution to eukaryotic biodiversity.

[Structure and Evolution of the AqE Gene in Insects].

The AqE gene encodes a sulfolactate dehydrogenase-like enzyme of the LDH2/MDG2 oxidoreductase family. The gene is found in bacteria and fungi, as well as in animals and plants whose lifestyles are associated with aquatic environments. The AqE gene is present in arthropods and, in particular, insects that are predominantly terrestrial. The distribution and structure of AqE was studied in insects in order to trace its evolutionary fate. The AqE gene was found to be absent from certain insect orders and suborders, being apparently lost. AqE duplication or multiplication was observed in some orders. AqE was found to vary both in length and intron-exon structure, from intronless to multi-intron. An ancient nature was demonstrated for AqE multiplication in insects, while younger duplications were also detected. It was assumed that a new function might be acquired by the gene with the formation of paralogs.

[Prevalence, Diversity, and Evolution of L18 (DD37E) Transposons in the Genomes of Cnidarians].

Transposable elements have a significant impact on the structure and functioning of multicellular genomes, and also serve as a source of new genes. Studying the diversity and evolution of transposable elements in different taxa is necessary for the fundamental understanding of their role in genomes. The Tc1/mariner elements are one of the most widespread and diverse groups of DNA transposons. In this work, the structure, distribution, diversity, and evolution of the L18 (DD37E) elements in the genomes of cnidarians (Cnidaria) were studied for the first time. As a result, it was found that the L18 group is an independent family (and not a subfamily of the TLE family, as previously thought) in the Tc1/mariner superfamily. Of the 51 detected elements, only four had potentially functional copies. It is assumed that the L18 transposons are of ancient origin, and, in addition, the elements found in the genomes of organisms of the Anthozoa and Hydrozoa classes do not come from a common ancestral transposon within the Cnidaria phylum. In organisms of the Hydrozoa class, L18 transposons appeared as a result of horizontal transfer at a later time period. An intraspecies comparison of the diversity of the L18 elements demonstrates high homogeneity with respect to "old" transposons, which have already lost their activity. At the same time, distant populations, as in the case of Hydra viridissima, have differences in the representation of DNA transposons and the number of copies. These data supplement the knowledge on the diversity and evolution of Tc1/mariner transposons and contribute to the study of the influence of mobile genetic elements on the evolution of multicellular organisms.

The Tc1-like elements with the spliceosomal introns in mollusk genomes.

Transposable elements (TEs) are DNA sequences capable of transpositions within the genome and thus exerting a considerable influence on the genome functioning and structure and serving as a source of new genes. TE biodiversity studies in previously unexplored species are important for the fundamental understanding of the TE influence on eukaryotic genomes. TEs are classified into retrotransposons and DNA transposons. IS630/Tc1/mariner (ITm) superfamily of DNA transposons is one of the most diverse groups broadly represented among the eukaryotes. The study of 19 mollusk genomes revealed a new group of ITm superfamily elements, which we henceforth refer to as TLEWI. These TEs are characterized by the low copy number, the lack of terminal inverted repeats, the catalytic domain with DD36E signature and the presence of spliceosomal introns in transposase coding sequence. Their prevalence among the mollusks is limited to the class Bivalvia. Since TLEWI possess the features of domesticated TE and structures similar to the eukaryotic genes which are not typical for the DNA transposons, we consider the hypothesis of co-optation of TLEWI gene by the bivalves. The results of our study will fill the gap of knowledge about the prevalence, activity, and evolution of the ITm DNA transposons in multicellular genomes and will facilitate our understanding of the mechanisms of TE domestication by the host genome.

Gene Encoding a Novel Enzyme of LDH2/MDH2 Family is Lost in Plant and Animal Genomes During Transition to Land.

L-Lactate/malate dehydrogenases (LDH/MDH) and type 2 L-lactate/malate dehydrogenases (LDH2/MDH2) belong to NADH/NADPH-dependent oxidoreductases (anaerobic dehydrogenases). They form a large protein superfamily with multiple enzyme homologs found in all branches of life: from bacteria and archaea to eukaryotes, and play an essential role in metabolism. Here, we describe the gene encoding a new enzyme of LDH2/MDH2 oxidoreductase family. This gene is found in genomes of all studied groups/classes of bacteria and fungi. In the plant kingdom, this gene was observed only in algae, but not in bryophyta or spermatophyta. This gene is present in all taxonomic groups of animal kingdom beginning with protozoa, but is lost in lungfishes and other, higher taxa of vertebrates (amphibians, reptiles, avians and mammals). Since the gene encoding the new enzyme is found only in taxa associated with the aquatic environment, we named it AqE (aquatic enzyme). We demonstrated that AqE gene is convergently lost in different independent lineages of animals and plants. Interestingly, the loss of the gene is consistently associated with transition from aquatic to terrestrial life forms, which suggests that this enzyme is essential in aquatic environment, but redundant or even detrimental in terrestrial organisms.

An Analysis of IS630/Tc1/mariner Transposons in the Genome of a Pacific Oyster, Crassostrea gigas.

Transposable elements represent the DNA fragments capable of increasing their copy number and moving within the genome. Class II mobile elements represents the DNA transposons, which transpose via excision and the subsequent reinsertion at random genomic loci. The increase of their copy number occurs only when the transposition event is coupled with the replication. IS630/Tc1/mariner DNA transposon superfamily is one of the largest and widely distributed among the Class II elements. In this work, we provide a detailed analysis of IS630/Tc1/mariner DNA transposons from the Pacific oyster, Crassostrea gigas. IS630/Tc1/mariner transposons represented in the genome of the Pacific oyster belong to four families, Tc1 (DD34E), mariner (DD34D), pogo (DDxD), and rosa (DD41D). More than a half of IS630/Tc1/mariner elements from C. gigas belong to Tc1 family. Furthermore, Mariner-31_CGi element was shown to represent a new and previously unknown family with DD37E signature. We also discovered the full-size transcripts of eight elements from Tc1, mariner, and pogo families, three of which can, presumably, retain their transposition activity.

[Analysis of expression of parental alleles Xist and Gla in interspecific embryonic hybrid cells during induced in vitro inactivation of X-chromosomes].

The results of in situ hybridization with labeled species specific and X-chromosome-specific probes suggest that hybrid cells obtained by fusion of Mus musculus embryonic stem cells (genotype XY) and splenocytes of M. caroli females contain two parental X-chromosomes. In five clones of hybrid cells, differentiation was induced in embryoid bodies in vitro, which was accompanied by inactivation of one of X-chromosomes. We analyzed the expression of Xist and Gla alleles in the embryoid bodies using RT-PCR with an account that expression of locus Xist is one of key events in X-chromosome inactivation, while gene Gla was used as a marker of active X-chromosome. Identification of allele transcripts of loci Xist and Gla was based on restriction polymorphism between M. musculus and M. caroli that we had described. Transcripts of both parental alleles of loci Xist and Gla were present in the embryoid bodies of all studied hybrid clones. No preferential inactivation of M. musculus or M. caroli X-chromosome was found in the tested embryonic hybrid cells despite the initial differences in ontogenetic status between X-chromosomes of embryonic stem cells and splenocytes.

["Chromosome memory" of parental genomes in embryonic hybrid cells]. / "Khromosomnaia pamiat'" roditel'skikh genomov v émbrional'nykh gibridnykh kletkakh.

In the hybrid cells obtained by fusion of embryonic stem cells with adult differentiated cells, homologous chromosomes are in two ontogenetic configurations: pluripotent and differentiated. In order to assess the role of cis- and trans-regulation in the maintenance of these states, we studied a set of clones of hybrid cells of the type embryonic stem cells-splenocytes and used two approaches: segregation of parental chromosomes and comparison of pluripotency of the past hybrid cells and embryonic stem cells. The segregation test showed that the hybrid cells lost only the homologs of the somatic partner and this process was sharply accelerated when the cells were cultivated in nonselective conditions, thus suggesting the full or partial preservation of the initial differences in the organization of parental homologs. The descendants of the former hybrid cells, which had the karyotype similar to that of embryonic stem cells, demonstrated the level of pluripotency, comparable with that of embryonic stem cells despite the long-term effect of trans-acting factors from the somatic partner in the genome of hybrid cells. The data obtained are interpreted in the framework of the concept of "chromosome memory", in the maintenance of which the key role is played by cis-regulatory factors.