Genome reshuffling of the copia element in an inbred line of Drosophila melanogaster

Biémont, Christian; Aouar, Ammaria; Arnault, C.

doi:10.1038/329742a0

Cited by 82 publications

(36 citation statements)

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“…We observed reversions of some previously fixed insertions in our experiment that might have caused partial rescue of fitness. Excisions of copia have been seen in one other experiment (Biemont et al, 1987). Excisions of other retrotransposons also seem to be rare (Belyaeva et al, 1982 ;Kim & Belyaeva, 1991;Nuzhdin & Mackay, 1994 ;Maside et al, 2000).…”

Section: (Iii) Tes In Mutation-accumulation Experimentsmentioning

confidence: 85%

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Mutation accumulation and the effect of copia insertions in Drosophila melanogaster

Houle

Nuzhdin

2004

Genet. Res.

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SummaryRepeated efforts to estimate the genomic deleterious mutation rate per generation (U) in Drosophila melanogaster have yielded inconsistent estimates ranging from 0 . 01 to nearly 1. We carried out a mutation-accumulation experiment with a cryopreserved control population in hopes of resolving some of the uncertainties raised by these estimates. Mutation accumulation (MA) was carried out by brother-sister mating of 150 sublines derived from two inbred lines. Fitness was measured under conditions chosen to mimic the ancestral laboratory environment of these genotypes. We monitored the insertions of a transposable element, copia, that proved to accumulate at the unusually high rate of 0 . 24 per genome per generation in one of our MA lines. Mutational variance in fitness increased at a rate consistent with previous studies, yielding a mutational coefficient of variation greater than 3 %. The performance of the cryopreserved control relative to the MA lines was inconsistent, so estimates of mutation rate by the Bateman-Mukai method are suspect. Taken at face value, these data suggest a modest decline in fitness of about 0 . 3 % per generation. The element number of copia was a significant predictor of fitness within generations ; on average, insertions caused a 0 . 76 % loss in fitness, although the confidence limits on this estimate are wide.

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Section: (Iii) Tes In Mutation-accumulation Experimentsmentioning

confidence: 85%

“…The rate of copia excision (owing to either precise or imprecise deletions) is interesting because of its rarity (Biemont et al, 1987;Pasyukova & Nuzhdin, 1993 ;Nuzhdin & Mackay, 1994). We detected the first copia excision from site 30A in generation 22 (no hybridization in six larvae of one MA replicate).…”

Section: (Iv) Transpositions Of Copia In Line 39mentioning

confidence: 85%

Mutation accumulation and the effect of copia insertions in Drosophila melanogaster

Houle

Nuzhdin

2004

Genet. Res.

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“…Finally, it should be noted that in three cases we found two additional neoRT copies in G418-resistant clones. Future evaluation of the significance of these unexpected structures will have to take into account that transposition (i) might be highly mutagenic for the retrotransposon as observed in yeast for Tyl (6) and (ii) under given circumstances might be a frequent and/or cooperative event as observed in Drosophila (17)(18)(19) …”

Section: Discussionmentioning

confidence: 99%

“…First, how can it be demonstrated that putative transposons actually transpose in mammalian cells; in particular, are proviral copies of the retrovirus able to transpose? Second, is transposition modulated by genetic or epigenetic factors as observed in Drosophila and yeast (6,(17)(18)(19)? Third, can transposition be implicated in tumorigenic processes?…”

Section: Introductionmentioning

confidence: 99%

An indicator gene to demonstrate intracellular transposition of defective retroviruses.

Heidmann

Nicolas

1988

Proc. Natl. Acad. Sci. U.S.A.

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An indicator gene for detection and quantitation of RNA-mediated transposition was constructed (neoRT). It was inserted into a Moloney murine leukemia provirus (Mo-MLV) (6). The homology between proviruses and "retrotransposons" (1, 6) is further strengthened by the occurrence of intracellular "virus-like particles" and of intermediates of reverse transcription in the case of copia and Tyl (7-11).In mammals, analysis of transposition is difficult to perform by using the biological assays that have been developed in Drosophila or yeast (6,12). Accordingly, putative transposons have been suspected from their structural similarities with proviruses and their high copy number in the genome. In the case of the IAP sequences, transposition was demonstrated by the discovery of their integration into unusual loci (13-15), a situation reminiscent of the retrovirus insertions observed in a number of tumors (16).These observations raise a series of questions. First, how can it be demonstrated that putative transposons actually transpose in mammalian cells; in particular, are proviral copies of the retrovirus able to transpose? Second, is transposition modulated by genetic or epigenetic factors as observed in Drosophila and yeast (6,(17)(18)(19)? Third, can transposition be implicated in tumorigenic processes?In addressing these questions, the potentially low frequency of transposition has to be taken into account (see refs. 20 and 21 for yeast and Drosophila). To analyze transposition in mammalian cells, we therefore constructed an "indicator gene for retrotransposition," which should allow the detection of transposition (by selective procedures) for any genetic element that transposes via an RNA intermediate. MATERIALS AND METHODSPlasmid Construction. To construct the retrotransposition indicator (neo)RT, the sequence for polyadenylylation of the thymidine kinase (tk) gene of the herpes simplex virus was isolated from plasmid pAGO (22) as a Sma I/Nco I 310-basepair (bp) fragment and was inserted at the unique BamHI site of pZip plasmid (23) between the Moloney murine leukemia virus (Mo-MLV) donor and acceptor splice sites after Klenow enzyme treatment of both fragments. The fragment containing the polyadenylylation sequence and the splice sites was next isolated as a Kpn I/Kpn I fragment and inserted between the SVtk promoter and the coding region of the neo gene at the Bgl II site of pSVtkneop3 (24) after Klenow treatment of both fragments; the neoRT indicator gene (Fig. L4) was then isolated as a HindIII/Sma I fragment.To delete the env genes in pMov3 (25) The final construction [pMo-MLV(neo)RTI was obtained upon ligation of the deleted pMov3 vector with the (neo)RT indicator gene after Klenow treatment (Fig. 1B).Cell Culture, Transfection, and Infection. Methods are described in ref. 27. Cells used were NIH 3T3 and FG10 (28), and the viral producer lines 3T3 Mo-MLV for Moloney and E404B for . 3T3 LacZ cells were a gift from D. Rocancourt and C. Bonnerot (Institut Pasteur). Selection of G418-resistant cells ...

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“…One surprising result was the observation of transposition bursts of TEs such as Doc (a non-LTR retrotransposon; Gerasimova et al 1990), copia (a retrotransposon; Biémont et al 1987;Gerasimova et al 1990;Pasyukova and Nuzhdin 1993), and P (a DNA transposon; Biémont et al 1990) while inbred lines were being maintained in the laboratory. Although we did not know whether this mobilization was the result of the inbreeding itself, or simply ''spontaneous'' transposition revealed by the homozygosity of the lines, which made it easier to observe new insertions, we were forced to conclude that inbred lines were not as homozygous as expected, at least with regard to TE insertions.…”

Section: Transposable Elements As Components Of Genetic Diversitymentioning

confidence: 99%

A Brief History of the Status of Transposable Elements: From Junk DNA to Major Players in Evolution

2010

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The idea that some genetic factors are able to move around chromosomes emerged more than 60 years ago when Barbara McClintock first suggested that such elements existed and had a major role in controlling gene expression and that they also have had a major influence in reshaping genomes in evolution. It was many years, however, before the accumulation of data and theories showed that this latter revolutionary idea was correct although, understandably, it fell far short of our present view of the significant influence of what are now known as ''transposable elements'' in evolution. In this article, I summarize the main events that influenced my thinking about transposable elements as a young scientist and the influence and role of these specific genomic elements in evolution over subsequent years. Today, we recognize that the findings about genomic changes affected by transposable elements have considerably altered our view of the ways in which genomes evolve and work.

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Genome reshuffling of the copia element in an inbred line of Drosophila melanogaster

Cited by 82 publications

References 19 publications

Mutation accumulation and the effect of copia insertions in Drosophila melanogaster

Mutation accumulation and the effect of copia insertions in Drosophila melanogaster

An indicator gene to demonstrate intracellular transposition of defective retroviruses.

A Brief History of the Status of Transposable Elements: From Junk DNA to Major Players in Evolution

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