P. G. DeBaryshe

Telomeres and telomerase: more than the end of the line.

Abstract. Early studies of telomerase suggested that telomeres are maintained by an elegant but relatively simple and highly conserved mechanism of telomerase-mediated 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.

Retrotransposons that maintain chromosome ends

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.

Evolution of diverse mechanisms for protecting chromosome ends by Drosophila TART telomere retrotransposons

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 (TARTvir) 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 (TARTmel) 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 TARTvir and HeT-Amel are essentially identical (using Tags), whereas HeT-Avir is clearly protected from end erosion by an as-yet-unspecified program.

Differential Maintenance of DNA Sequences in Telomeric and Centromeric Heterochromatin

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.

Evolution of species-specific promoter-associated mechanisms for protecting chromosome ends by Drosophila Het-A telomeric transposons.

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.

Another protozoan contributes to understanding telomeres and transposable elements

Transposable elements (TEs) are abundant in the genomes of eukaryotes, making up >40% of the nuclear DNA of humans (1). Many TEs appear to be simply selfish DNA “whose primary and often only function is self-preservation” (2). Of course, selfish behavior has effects on bystanders, and TEs can have a number of effects on the rest of the genome. TEs can affect gene expression by inserting within a gene and disrupting its structure, or by directing transcription of a nearby gene. TEs also can affect splicing and transcription termination, thereby altering the protein product of a gene. TEs can insert at multiple sites in the genome, allowing recombination between otherwise nonhomologous regions, resulting in large-scale rearrangements of chromosomes. In fact, many of the best-known TEs were initially identified because they mutated a gene under study. TEs can even have incidental benefits for the host, and there is one example of TEs devoted entirely to the benefit of the genome in which they reside, the Drosophila telomeric transposons, HeT-A and TART. In this issue of PNAS, Arkhipova and Morrison (3) report two very interesting TEs from Giardia lamblia that may also have evolved to benefit their host. This study adds another piece to the emerging picture of what happens at chromosome ends.