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Characterization of a highly conserved sequence related to mutator transposable elements in maize.

Mutator stocks of maize exhibit a high mutation rate correlated with the activity of a family of transposable elements. Mu1 and, to a lesser extent, the closely related Mu1.7 elements are responsible for most mutator-induced mutations that have been characterized. These elements are found in 10-60 copies in mutator stocks, and zero to a few intact elements exist in nonmutator maize stocks. Additionally, the component parts of Mu elements exist separately in the maize genome. The Mu terminal inverted repeats are found in multiple copies in all maize lines and related Zea species tested, and Mu internal sequences exist unassociated with Mu termini. In the present paper, we describe the structure and genomic distribution of one Mu-homologous sequence termed MRS-A (for Mu-related sequence). DNA sequencing shows that MRS-A is closely related to the internal region of Mu1 and Mu1.7 elements. However, it has no Mu termini and does not have the structure of a transposable element. This sequence is present in one or two copies in all maize lines and is highly conserved in the genus Zea. A similar sequence exists in a species within the genus most closely related to Zea, Tripsacum dactyloides, although the T. dactyloides genome does not contain any Mu termini or intact Mu elements. Furthermore, an RNA transcript homologous to MRS-A and its flanking DNA is found in both mutator and nonmutator maize plants. These results suggest that MRS-A represents a stable, functional region of the maize genome, and we speculate that a similar sequence was encompassed by Mu termini to generate a Mu transposable element.

Base Sequence↗

The Adaptive Roles of Active Transposable Elements in Insect Hosts.

Active transposable elements (TEs) are capable of generating new insertions in genomes and have historically been viewed as genomic parasites due to their largely detrimental or neutral effects. However, emerging evidence suggests that these elements also play a crucial role in driving adaptive evolution in insects. This mini-review synthesizes recent findings on how active TEs contribute to insect adaptation through various mechanisms, including regulation of gene expression, structural variation, and epigenetic effects. Notable examples of adaptation driven by active TEs include their roles in insecticide resistance, morphological adaptations, tolerance to harsh climates, and antiviral immune responses. We argue that while host silencing mechanisms, such as the piRNA pathway, tightly regulate TE activity to minimize harmful effects, the context-dependent activation of active TEs can generate beneficial genetic variation that enhances insect adaptations to anthropogenic and climatic pressures. Future research that integrates long-read sequencing, single-cell omics, and gene editing techniques will provide a robust mechanistic foundation for understanding the adaptive significance of active TEs in insects, with important implications for pest management, pollinator protection, and evolutionary biology.

Journal Article↗

Non-random distribution of transposable elements in the nuclear genome of plants.

We have studied the genomic distribution of five different families of plant transposable elements by analyzing their location in DNA fractions from maize and tobacco genomes fractionated according to base composition. The results show that each family of elements is preferentially integrated in one specific fraction of its respective host genome. This demonstrates that the distribution of transposable elements in the nuclear genome of plants is not random but compartmentalized, i.e., the elements are located in specific genomic compartments characterized by having a specific G+C content and representing a small proportion of the genomes. Furthermore, these compartments seem to correspond to the genomic regions where most of the plant genes are also located, suggesting a preferential integration of transposable elements in the transcriptionally active regions of the plant genome. The implications of these results on the current applications of transposon tagging techniques are discussed.

Base Composition↗

Identification of putative nonautonomous transposable elements associated with several transposon families in Caenorhabditis elegans.

Putative nonautonomous transposable elements related to the autonomous transposons Tc1, Tc2, Tc5, and mariner were identified in the C. elegans database by computational analysis. These elements are found throughout the C. elegans genome and are defined by terminal inverted repeats with regions of sequence similarity, or identity, to the autonomous transposons. Similarity between loci containing related nonautonomous elements ends at, or near, the boundaries of the terminal inverted repeats. In most cases the terminal inverted repeats of the putative nonautonomous transposable elements are flanked by potential target-site duplications consistent with the associated autonomous elements. The nonautonomous elements identified vary considerably in size (from 100 bp to 1.5 kb in length) and copy number in the available database and are localized to introns and flanking regions of a wide variety of C. elegans genes.

Animals↗

I transposable elements and I-R hybrid dysgenesis in Drosophila.

I factors, transposable elements related to mammalian LINEs, are responsible for I-R hybrid dysgenesis in Drosophila melanogaster. Although they are not structurally related to retrovirus-like transposable elements, they appear to move around the genome via reverse transcription of a full-length RNA intermediate. The mechanism and control of this process are now being dissected at the molecular level.

Animals↗

The behavior of a Daphnia pulex transposable element in cyclically and obligately parthenogenetic populations.

Using Southern blot analysis, we have characterized restriction fragment patterns of a transposable element. Pokey, in obligately and cyclically parthenogenetic populations of the cladoceran crustacean Daphnia pulex. We show that the element is most likely active in cyclically parthenogenetic populations but is, for the most part, inactive in obligate parthenogens. This result is consistent with theory suggesting that transposable element dynamics are likely to change with a change in reproductive mode. Such changes could have important consequences for the long-term evolutionary potential of obligate parthenogens and may also be informative with regard to the underlying mechanisms that regulate transposable element frequencies in sexual organisms.

Animals↗

Tnt1, a mobile retroviral-like transposable element of tobacco isolated by plant cell genetics.

Transposable elements can be identified by their ability to induce mutant alleles at new loci. The retrotransposon family is thought to transpose through an RNA intermediate and has many similarities to vertebrate proretroviruses. In plants, retrotransposons have been described in maize, Arabidopsis and wheat, and non-viral retroposons in maize. Most of these elements, however, have been found as non-mobile integrated units. Here, we report the isolation of the first tobacco (Nicotiana tabacum) transposable element, Tnt1, which seems to be the most complete mobile retrotransposon characterized in higher plants. Tnt1 has been isolated after its transposition into the nitrate reductase (NR) structural gene of tobacco, and transposition events have been detected through in vitro selection of spontaneous NR-deficient (NR-) mutant lines in cell cultures derived from tobacco mesophyll protoplasts. Tnt1 is 5,334 nucleotides long, contains two 610-base-pair-long terminal repeats and a single open reading frame of 3,984 nucleotides. Comparison of the Tnt1 open reading frame coding potential with those of the Drosophila melanogaster copia retrotransposon, yeast Ty retrotransposon, and vertebrate proretroviruses revealed that Tnt1 is closely related to copia and carries all the functions known to be required for autonomous transposition by reverse transcription.

Alleles↗

Spontaneous changes in Drosophila melanogaster transposable elements and their effects on fitness.

Twenty-eight spontaneous alterations modifying the hybridization banding pattern of six families of transposable elements (297, Foldback, copia, jockey, P and hobo) have been fixed in a set of mutation-accumulation lines of Drosophila melanogaster. Their effect on fitness has been studied by competition with the original pattern. Most alterations affecting transposable elements were shown to be rearrangements with no detectable effect on fitness, showing that spontaneous transposable element mutations mainly generate minor fitness mutations.

Animals↗

Genome-wide distribution and potential regulatory functions of AtATE, a novel family of miniature inverted-repeat transposable elements in Arabidopsis thaliana.

A study of transgenic promoter::beta-glucuronidase lines showed that the promoters of the two Arabidopsis ARGININE DECARBOXYLASE paralogues, ADC1 and ADC2, exhibited extremely different patterns of activity. One major feature of the promoter of ADC1 was the presence of a novel transposable element, which was shown to possess all of the characteristics of Miniature Inverted-repeat Transposable Elements (MITEs), and to be present in 26 full-length copies and 1617 partial copies and fragments distributed throughout the Arabidopsis genome. TRANSFAC analysis showed that this transposable element possesses a significant number of transcription-factor binding motifs. A bioinformatics approach based on a suffix-tree compilation was used to obtain an exhaustive description of exact copy numbers and positions of the element in the Arabidopsis genome. The distribution among the chromosomes was non-random, and a significant number of copies were found in regions flanking genes. Full-length copies of the transposable element were detected in the immediate vicinity of 22 genes, either upstream or downstream.

Arabidopsis↗

Target sequences for the C. elegans transposable element Tc1.

The target sequences for two independent insertions of the transposable element Tc1 from Caenorhabditis elegans show homology. Because both insertions are at palindromic TA/AT sequences, the exact boundaries of Tc1 cannot be distinguished; Tc1 could be 1610 bp and flanked by a 2-bp duplication of the target site or it could be 1612 bp and without target site duplication. The latter possibility implies a novel manner for insertion of a transposable element.

Animals↗

Formation and loss of large, unstable tandem arrays of the piggyBac transposable element in the yellow fever mosquito, Aedes aegypti.

The Class II transposable element, piggyBac, was used to transform the yellow fever mosquito, Aedes aegypti. In two transformed lines only 15-30% of progeny inherited the transgene, with these individuals displaying mosaic expression of the EGFP marker gene. Southern analyses, gene amplification of genomic DNA, and plasmid rescue experiments provided evidence that these lines contained a high copy number of piggyBac transformation constructs and that much of this DNA consisted of both donor and helper plasmids. A detailed analysis of one line showed that the majority of piggyBac sequences were unit-length donor or helper plasmids arranged in a large tandem array that could be lost en masse in a single generation. Despite the presence of a transposase source and many intact donor elements, no conservative (cut and paste) transposition of piggyBac was observed in these lines. These results reveal one possible outcome of uncontrolled and/or unexpected recombination in this mosquito, and support the conclusion that further investigation is necessary before transposable elements such as piggyBac can be used as genetic drive mechanisms to move pathogen-resistance genes into mosquito populations.

Aedes↗

The transposable element mariner mediates germline transformation in Drosophila melanogaster.

A vector for germline transformation in Drosophila melanogaster was constructed using the transposable element mariner. The vector, denoted pMlwB, contains a mariner element disrupted by an insertion containing the wild-type white gene from D. melanogaster, the beta-galactosidase gene from Escherichia coli and sequences that enable plasmid replication and selection in E. coli. The white gene is controlled by the promoter of the D. melanogaster gene for heat-shock protein 70, and the beta-galactosidase gene is flanked upstream by the promoter of the transposable element P as well as that of mariner. The MlwB element was introduced into the germline of D. melanogaster by co-injection into embryos with an active mariner element, Mos1, which codes for a functional transposase and serves as a helper. Two independent germline insertions were isolated and characterized. The results show that the MlwB element inserted into the genome in a mariner-dependent manner with the termini of the inverted repeats inserted at a TA dinucleotide. Both insertions exhibit an unexpected degree of germline and somatic stability, even in the presence of an active mariner element in the genetic background. These results demonstrate that the mariner transposable element, which is small (1286 bp) and relatively homogeneous in size among different copies, is nevertheless capable of promoting the insertion of the large (13.2 kb) MlwB element. Because of the widespread phylogenetic distribution of mariner among insects, these results suggest that mariner might provide a wide host-range transformation vector for insects.

Animals↗

Sequence of the C. elegans transposable element Tc1.

The complete nucleotide sequence was determined for Tc1, a transposable element in the nematode Caenorhabditis elegans. The 1610-base-pair element terminates in 54-base-pair perfect inverted repeats and is flanked by a 2-base-pair duplication of the target sequence. The Tc1 sequence contains two long open reading frames on the same DNA strand but in different translational reading frames. The positions of transcriptional control sequences suggest that a single transcript is made, which could produce two polypeptides, 273 and 112 amino acids in length. These features, i.e. terminal repeats, target site duplication and open reading frames, make Tc1 similar to transposable elements from other species.

Animals↗

The role of host factors in the population dynamics of selfish transposable elements.

Previous models of the evolution of selfish transposable genetic elements have failed to include the possibility that transposition may be limited by shortage of a host-encoded factor. The titration of host factors may be important in limiting the rate of transpositional increase in these elements. This will be exacerbated if multiple copies of the host factor protein must bind simultaneously to the target element. In the case of the Drosophila melanogaster P transposable element, which can exist as autonomous and as non-autonomous copies, there is evidence that a host-encoded protein, IRBP, is required for the transposition process. We have produced a specific model of the invasion of a host population by the P element, in which we have incorporated the requirement for the multiple binding of a host factor. We find that, for the P family, in which it is apparently transposition itself that creates selective harm to the host, the effect of selection in the context of host factor limitation is to drive up copy number. This can result in a novel high copy number-low transposition state. We also find that host factor limitation reinforces the tendency for transposable elements that create sterility to be replaced by their deletion derivatives.

Animals↗

Transposable elements as initiators of insecticide resistance.

Genetic change that leads to insecticide resistance is poorly understood for most resistance genes. Transposable elements have been shown to be powerful mutagenic agents in Drosophila melanogaster (Meigen), and they potentially can create genetic change leading to resistance in pest insect populations. Here the characteristic of transposable elements and the consequences of their movement in D. melanogaster and other insects are discussed, and reports of their involvement in insecticide resistance are examined. I conclude that transposable elements must be considered as mutagenic agents that can lead to resistance. Their involvement needs to be directly addressed in studies of resistant insects, particularly by investigators involved in cloning resistance genes.

Animals↗

Integration target site selection for retroviruses and transposable elements.

When a retrovirus infects a cell, its RNA genome is reverse transcribed into a double-stranded DNA, which is then permanently integrated into the host chromosome. Integration is one of the essential steps in the retroviral life cycle. Many transposable elements also move around and integrate into the host genome as part of their life cycle, some through RNA intermediates and some through 'cut and paste' mechanisms. Integration of retroviruses and transposable elements into 'sensitive areas' of the genome can cause irreparable damage. On the other hand, because of their ability to integrate permanently, and the relatively efficient rates of transgenesis, retroviruses and transposable elements are widely used as gene delivery tools in basic research and gene therapy trials. Recent events in gene therapy treatments for X-linked severe combined immunity deficiencies (X-SCID) have highlighted both the promise and some of the risks involved with utilizing retroviruses. Nine of 11 children were successfully treated for X-SCID using a retrovirus carrying the gene mutated in this disease. However, later two of these children developed leukemias because of retroviral integrations in the putative oncogene LMO2 [1]. A third child has also been demonstrated to have an integration in LMO2, but is as of yet nonsymptomatic [2]. It is a bit difficult to explain the high frequency of integrations into the same gene using a random model of retroviral integration, and there has been evidence for decades that retroviral integrations may not be random. But the data were somewhat limited in their power to determine the precise nature of the integration biases. The completion of the human genome sequence coupled with sensitive polymerase chain reaction techniques and an ever-decreasing cost of sequencing has given a powerful new tool to the study of integration site selection. In this review, we describe the findings from several recent global surveys of target site selection by retroviruses and transposable elements, and discuss the possible ramifications of these findings to both mechanisms of action and to the use of these elements as gene therapy vectors.

Animals↗

Analysis of a mutator activity necessary for germline transposition and excision of Tc1 transposable elements in Caenorhabditis elegans.

The Tc1 transposable element family of the nematode Caenorhabditis elegans consists primarily of 1.6-kb size elements. This uniformity of size is in contrast to P in Drosophila and Ac/Ds in maize. Germline transposition and excision of Tc1 are detectable in the Bergerac (BO) strain, but not in the commonly used Bristol (N2) strain. A previous study suggested that multiple genetic components are responsible for the germline Tc1 activity of the BO strain. To analyze further this mutator activity, we derived hybrid strains between the BO strain and the N2 strain. One of the hybrid strains exhibits a single locus of mutator activity, designated mut-4, which maps to LGI. Two additional mutators, mut-5 II and mut-6 IV, arose spontaneously in mut-4 harboring strains. This spontaneous appearance of mutator activity at new sites suggests that the mutator itself transposes. The single mutator-harboring strains with low Tc1 copy number generated in this study should be useful in investigations of the molecular basis of mutator activity. As a first step toward this goal, we examined the Tc1 elements in these low copy number strains for elements consistently co-segregating with mutator activity. Three possible candidates were identified: none was larger than 1.6 kb.

Animals↗

Properties of transgenic strains of Drosophila melanogaster containing I transposable elements from Drosophila teissieri.

I factors are transposable elements of Drosophila melanogaster similar to mammalian LINEs, that transpose by reverse transcription of an RNA intermediate and are responsible for the I-R system of hybrid dysgenesis. There are two categories of strains in this species: inducer, that contain about 15 I elements at the various sites on chromosomal arms, and reactive, that lack active I factors. I elements occur in various Drosophila species. Potentially functional I factors from Drosophila teissieri can transpose when introduced by P-element-mediated transformation in a reactive strain of Drosophila melanogaster. We have studied the properties of Drosophila melanogaster strains into which such an I factor from Drosophila teissieri, named Itei, was introduced. Typical hybrid dysgenesis is produced when males carrying Itei are crossed with reactive females. However, more than one copy of the element seems necessary to produce dysgenic traits, whereas only one I factor of Drosophila melanogaster seems to be sufficient. The copy number of Itei in transformed lines maintained by endogamous crosses increases rapidly and stabilizes at values similar to those observed in inducer strains. As Drosophila teissieri contains much fewer copies than the Drosophila melanogaster strains, this suggests that the copy number of I elements is not simply regulated by sequences present in the element itself.

Animals↗