RESUMEN
Reverse transcriptases have shaped genomes in many ways. A remarkable example of this shaping is found on telomeres of the genus Drosophila, where retrotransposons have a vital role in chromosome structure. Drosophila lacks telomerase; instead, three telomere-specific retrotransposons maintain chromosome ends. Repeated transpositions to chromosome ends produce long head to tail arrays of these elements. In both form and function, these arrays are analogous to the arrays of repeats added by telomerase to chromosomes in other organisms. Distantly related Drosophila exhibit this variant mechanism of telomere maintenance, which was established before the separation of extant Drosophila species. Nevertheless, the telomere-specific elements still have the hallmarks that characterize non-long terminal repeat (non-LTR) retrotransposons; they have also acquired characteristics associated with their roles at telomeres. These telomeric retrotransposons have shaped the Drosophila genome, but they have also been shaped by the genome. Here, we discuss ways in which these three telomere-specific retrotransposons have been modified for their roles in Drosophila chromosomes.
Asunto(s)
Cromosomas/ultraestructura , Retroelementos/genética , Animales , Centrómero/ultraestructura , Drosophila/genética , Drosophila melanogaster , Eucromatina/química , Eliminación de Gen , Heterocromatina/química , Modelos Genéticos , Filogenia , ARN sin Sentido/genética , Telómero/ultraestructura , Cromosoma Y/ultraestructuraRESUMEN
Repeated DNA in heterochromatin presents enormous difficulties for whole-genome sequencing; hence, sequence organization in a significant portion of the genomes of multicellular organisms is relatively unknown. Two sequenced BACs now allow us to compare telomeric retrotransposon arrays from Drosophila melanogaster telomeres with an array of telomeric retrotransposons that transposed into the centromeric region of the Y chromosome >13 MYA, providing a unique opportunity to compare the structural evolution of this retrotransposon in two contexts. We find that these retrotransposon arrays, both heterochromatic, are maintained quite differently, resulting in sequence organizations that apparently reflect different roles in the two chromosomal environments. The telomere array has grown only by transposition of new elements to the chromosome end; the centromeric array instead has grown by repeated amplifications of segments of the original telomere array. Many elements in the telomere have been variably 5'-truncated apparently by gradual erosion and irregular deletions of the chromosome end; however, a significant fraction (4 and possibly 5 or 6 of 15 elements examined) remain complete and capable of further retrotransposition. In contrast, each element in the centromere region has lost ≥ 40% of its sequence by internal, rather than terminal, deletions, and no element retains a significant part of the original coding region. Thus the centromeric array has been restructured to resemble the highly repetitive satellite sequences typical of centromeres in multicellular organisms, whereas, over a similar or longer time period, the telomere array has maintained its ability to provide retrotransposons competent to extend telomere ends.
Asunto(s)
Centrómero/genética , ADN/genética , Drosophila melanogaster/genética , Heterocromatina/genética , Telómero/genética , Animales , Drosophila melanogaster/citología , Heterocromatina/metabolismo , Retroelementos/genética , Análisis de Secuencia de ADN , Eliminación de SecuenciaRESUMEN
The retrotransposons HeT-A, TART, and TAHRE, which maintain Drosophila telomeres, transpose specifically onto chromosome ends to form long arrays that extend the chromosome and compensate for terminal loss. Because they transpose by target-primed reverse transcription, each element is oriented so that its 5' end serves as the extreme end of the chromosome until another element transposes to occupy the terminal position. Thus 5' sequences are at risk for terminal erosion while the element is at the chromosome end. Here we report that TART elements in Drosophila melanogaster and Drosophila virilis show species-specific innovations in promoter architecture that buffer loss of sequence exposed at chromosome ends. The two elements have evolved different ways to effect this protection. The D. virilis TART (TART(vir)) promoter is found in the 3' UTR of the element directly upstream of the element transcribed. Transcription starts within the upstream element so that a "Tag" of extra sequence is added to the 5' end of the newly transcribed RNA. This Tag provides expendable sequence to buffer end erosion of essential 5' sequence after the RNA is reverse transcribed onto the chromosome. In contrast, the D. melanogaster TART (TART(mel)) promoter initiates transcription deep within the 5' UTR, but the element is able to replace and extend the 5' UTR sequence by copying sequence from its 3' UTR, we believe while being reverse transcribed onto the chromosome end. Astonishingly, end-protection in TART(vir) and HeT-A(mel) are essentially identical (using Tags), whereas HeT-A(vir) is clearly protected from end erosion by an as-yet-unspecified program.
Asunto(s)
Región de Flanqueo 5'/genética , Cromosomas , Proteínas de Drosophila/genética , Drosophila/genética , Retroelementos/genética , Telómero/genética , Animales , Evolución Biológica , Drosophila melanogaster/genética , Regiones Promotoras Genéticas , Transcripción GenéticaRESUMEN
The non-LTR retrotransposons forming Drosophila telomeres constitute a robust mechanism for telomere maintenance, one which has persisted since before separation of the extant Drosophila species. These elements in D. melanogaster differ from nontelomeric retrotransposons in ways that give insight into general telomere biology. Here, we analyze telomere-specific retrotransposons from D. virilis, separated from D. melanogaster by 40 to 60 million years, to evaluate the evolutionary divergence of their telomeric traits. The telomeric retrotransposon HeT-A from D. melanogaster has an unusual promoter near its 3' terminus that drives not the element in which it resides, but the adjacent downstream element in a head-to-tail array. An obvious benefit of this promoter is that it adds nonessential sequence to the 5' end of each transcript, which is reverse transcribed and added to the chromosome. Because the 5' end of each newly transposed element forms the end of the chromosome until another element transposes onto it, this nonessential sequence can buffer erosion of sequence essential for HeT-A. Surprisingly, we have now found that HeT-A in D. virilis has a promoter typical of non-LTR retrotransposons. This promoter adds no buffering sequence; nevertheless, the complete 5' end of the element persists in telomere arrays, necessitating a more precise processing of the extreme end of the telomere in D. virilis.
Asunto(s)
Elementos Transponibles de ADN/genética , Drosophila/genética , Evolución Molecular , Regiones Promotoras Genéticas/genética , Telómero/genética , Regiones no Traducidas 3'/genética , Regiones no Traducidas 5'/genética , Animales , Secuencia de Bases , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , ARN sin Sentido/genética , Homología de Secuencia de Ácido Nucleico , Especificidad de la Especie , Transcripción GenéticaRESUMEN
Telomeres across the genus Drosophila are maintained, not by telomerase, but by two non-LTR retrotransposons, HeT-A and TART, that transpose specifically to chromosome ends. Successive transpositions result in long head-to-tail arrays of these elements. Thus Drosophila telomeres, like those produced by telomerase, consist of repeated sequences reverse transcribed from RNA templates. The Drosophila repeats, complete and 5'-truncated copies of HeT-A and TART, are more complex than telomerase repeats; nevertheless, these evolutionary variants have functional similarities to the more common telomeres. Like other telomeres, the Drosophila arrays are dynamic, fluctuating around an average length that can be changed by changes in the genetic background. Several proteins that interact with telomeres in other species have been found to have homologues that interact with Drosophila telomeres. Although they have hallmarks of non-LTR retrotransposons, HeT-A and TART appear to have a special relationship to Drosophila. Their Gag proteins are efficiently transported into diploid nuclei where HeT-A Gag recruits TART Gag to chromosome ends. Gags of other non-LTR elements remain predominantly in the cytoplasm. These studies provide intriguing evolutionary links between telomeres and retrotransposable elements.
Asunto(s)
Drosophila/genética , Retroelementos/genética , Telómero/genética , Animales , Núcleo Celular/genética , Cromosomas/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Evolución Molecular , Productos del Gen gag/genética , Productos del Gen gag/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Microscopía Fluorescente , Estructura Terciaria de Proteína/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Retroelementos/fisiología , Telómero/metabolismoRESUMEN
Telomere molecular biology is far more complex than originally thought. Understanding biological systems is aided by study of evolutionary variants, and Drosophila telomeres are remarkable variants. Drosophila lack telomerase and the arrays of simple repeats generated by telomerase in almost all other organisms; instead, Drosophila telomeres are long tandem arrays of two non-LTR retrotransposons, HeT-A and TART. These are the first transposable elements found to have a bona fide role in cell structure, revealing an unexpected link between telomeres and what is generally considered to be parasitic DNA. In addition to providing insight into the cellular functions performed by telomeres, analysis of HeT-A and TART is providing insight into the evolution of chromosomes, retrotransposons, and retroviruses. Recent studies show that retrotransposon telomeres constitute a robust system for maintaining chromosome ends. These telomeres are now known to predate the separation of extant Drosophila species, allowing ample time for elements and hosts to coevolve interesting mechanisms.
Asunto(s)
Evolución Biológica , Retroelementos/fisiología , Telómero/fisiología , Animales , Drosophila/genética , Drosophila/fisiología , Proteínas de Drosophila/fisiología , Productos del Gen gag/fisiología , Telomerasa/fisiologíaRESUMEN
In Drosophila two non-LTR retrotransposons, HeT-A and TART, offer a novel experimental system for the study of heterochromatin. These elements, found only in heterochromatin, form Drosophila telomeres by repeated transposition onto chromosome ends. Their transposition yields arrays of repeats larger and more irregular than the repeats produced by telomerase; nevertheless, the transpositions are, in principle, equivalent to the telomere-building action of telomerase. The identification of the HeT-A promoter has given the first view of the molecular structure of a promoter active in heterochromatin. These telomere-specific elements are unusual in having a large amount of non-coding sequence. Like many other heterochromatic sequences, the HeT-A non-coding sequence has a repetitive organization strongly conserved within the species, although the sequence itself can undergo significant change between species (a typical example of concerted evolution). Such heterochromatic sequences could be important for the cell, perhaps as docking stations for essential proteins.
Asunto(s)
Elementos Transponibles de ADN , Drosophila/genética , Heterocromatina/genética , Telómero , Animales , Regiones Promotoras GenéticasRESUMEN
Early studies of telomerase suggested that telomeres are maintained by an elegant but relatively simple and highly conserved mechanism of telomerase-medicated replication. As we learn more, it has become clear that the mechanism is elegant but not as simple as first thought. It is also evident that, although many species use similar, sometimes identical, DNA sequences for telomeres, these species express their own individuality in the way they regulate these sequences and, perhaps, in the additional tasks that they have imposed on their telomeric DNA. The striking similarities between telomeres in different species have revealed much about chromosome ends; the differences are proving to be equally informative. In addition to the differences between species that use telomerase, there are also a few exceptional organisms with atypical telomeres for which no telomerase activity has been detected. This review addresses recent studies, the insights they offer, and, perhaps more importantly, the questions they raise.
Asunto(s)
Telomerasa/metabolismo , Telómero , Envejecimiento/genética , Animales , Humanos , Neoplasias/genéticaRESUMEN
Telomeres in Drosophila melanogaster are composed of multiple copies of two retrotransposable elements, HeT-A and TART instead of the short DNA repeats generated by telomerase in most organisms. Transpositions of HeT-A and TART yield arrays of repeats larger and more irregular than the repeats produced by telomerase; nevertheless, these transpositions are, in principle, equivalent to the telomere-building action of telomerase. Both telomerase and transposition of HeT-A and TART extend chromosomes by RNA-templated addition of specific sequences. We have proposed that HeT-A has evolved from genes encoding telomerase components. Although both HeT-A and TART share some novel features, TART probably has a different origin from HeT-A. HeT-A and TART are clearly identifiable as non-long terminal repeat (non-LTR) retrotransposons. Both telomere elements transpose only to the ends of chromosomes (apparently to any chromosome end in D. melanogaster) and each contains a large segment of untranslated sequence. HeT-A and TART are the first examples of transposable elements with a clear role in chromosome structure. This has interesting implications for the evolution of both chromosomes and transposable elements. The finding also raises the possibility that other transposable elements with bona fide roles in the cell will be detected, not only in Drosophila, but also in other organisms.
Asunto(s)
Cromosomas , Elementos Transponibles de ADN , Drosophila melanogaster/genética , Telómero , Animales , Evolución Molecular , Regiones Promotoras Genéticas , Retroviridae/genéticaRESUMEN
The transposable elements HeT-A and TART constitute the telomeres of Drosophila chromosomes. Both are non-long terminal repeat (LTR) retrotransposons, sharing the remarkable property of transposing only to chromosome ends. In addition, strong sequence similarity of their gag proteins indicates that these coding regions share a common ancestor. These findings led to the assumption that HeT-A and TART are closely related. However, we now find that these elements produce quite different sets of transcripts. HeT-A produces only sense-strand transcripts of the full-length element, whereas TART produces both sense and antisense full-length RNAs, with antisense transcripts in more than 10-fold excess over sense RNA. In addition, features of TART sequence organization resemble those of a subclass of non-LTR elements characterized by unequal terminal repeats. Thus, the ancestral gag sequence appears to have become incorporated in two different types of elements, possibly with different functions in the telomere. HeT-A transcripts are found in both nuclear and cytoplasmic cell fractions, consistent with roles as both mRNA and transposition template. In contrast, both sense and antisense TART transcripts are almost entirely concentrated in nuclear fractions. Also, TART open reading frame 2 probes detect a cytoplasmic mRNA for reverse transcriptase (RT), with no similarity to TART sequence 5' or 3' of the RT coding region. This RNA could be a processed TART transcript or the product of a "free-standing" RT gene. Either origin would be novel. The distinctive transcription patterns of both HeT-A and TART are conserved in Drosophila yakuba, despite significant sequence divergence. The conservation argues that these sets of transcripts are important to the function(s) of HeT-A and TART.