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A J Flavell

Publications and source records attributed to A J Flavell.

At least 19 recordsLinked to original sources
PubMed1995

Ty1-copia group retrotransposon sequences in amphibia and reptilia.

We have isolated sequences belonging to Ty1-copia group retrotransposons from the genomes of an amphibian (Pyxicephalus adspersa) and three reptiles (Conolophus subscristatus, Amblyrynchus cristatus and Pytas mucosus). Two different sequences were found in the amphibian (Tpa1 and Tpa2). Each is present in several copies per genome and absent from the genomes of two other amphibian species. The C. subcristatus sequence Tcs1 is present in multiple copies in both its host genome (Galapagos land iguana) and the genome of the related Galapagos marine iguana (A. cristatus). There is little or no polymorphism in Tcs1 insertions between different individual animals, suggesting that this sequence is not transposing rapidly in either iguana genome. The P. mucosus sequence Tpm1 shows a discontinuous distribution in snake species, suggesting that it has either been lost from many lineages during vertical germline transmission or has been transferred horizontally in some snake species. Phylogenetic comparisons of all these sequences with each other and with other members of this retrotransposon group from other animals and plants show that sequences within a particular vertebrate species are most closely related to each other, consistent with a vertical transmission model for their evolution.

Amino Acid Sequence↗
PubMed1995

Retroelements, reverse transcriptase and evolution.

Retroelements are genetic elements that can exist as DNA or RNA or DNA/RNA duplexes. Although retroviruses are the best known retroelements, there are many other types, including close relatives of retroviruses like LTR retrotransposons, more distant relatives like non-LTR retrotransposons, caulimoviruses and hepadnaviruses and elements with virtually no similarity, like retrons. Virtually all retroelements are 'selfish DNAs' with no involvement with the normal development or maintenance of their host cells, the only known exception being telomereres/telomerases which maintain the ends of chromosomes. Virtually all retroelements use tRNA, or RNA with strong secondary structure, to initiate their reverse transcription. The coincidence between the use of tRNA, a molecule central to the conversion of RNA to protein, with reverse transcriptase, an enzyme which is crucial for the conversion of RNA to DNA is striking, because RNA probably preceded DNA and protein in evolution. It seems plausible that retroelements were present at the genesis of living systems.

Base Sequence↗
PubMed1992

Ty1-copia group retrotransposons are ubiquitous and heterogeneous in higher plants.

We have used the polymerase chain reaction to isolate fragments of Ty1-copia group retrotransposons from a wide variety of members of the higher plant kingdom. 56 out of 57 species tested generate an amplified fragment of the size expected for reverse transcriptase fragments of Ty1-copia group retrotransposons. Sequence analysis of subclones shows that the PCR fragments display varying degrees of sequence heterogeneity. Sequence heterogeneity therefore seems a general property of Ty1-copia group retrotransposons of higher plants, in contrast to the limited diversity seen in retrotransposons of Saccharomyces cerevisiae and Drosophila melanogaster. Phylogenetic analysis of all these sequences shows, with some significant exceptions, that the degree of sequence divergence in the retrotransposon populations between any pair of species is proportional to the evolutionary distance between those species. This implies that sequence divergence during vertical transmission of Ty1-copia group retrotransposons within plant lineages has been a major factor in the evolution of Ty1-copia group retrotransposons in higher plants. Additionally, we suggest that horizontal transmission of this transposon group between different species has also played a role in this process.

Amino Acid Sequence↗
PubMed1992

Expression and sequence analysis of cDNAs induced during the early stages of tuberisation in different organs of the potato plant (Solanum tuberosum L.).

cDNA clones of two genes (TUB8 and TUB13) which show a 25-30-fold increase in transcript in the stolon tip during the early stages of tuberisation, have been isolated by differential screening. These genes are also expressed in leaves, stems and roots and the expression pattern in these organs changes on tuberisation. Southern analysis shows homologous sequences in the non-tuberising wild type potato species Solanum brevidens and in Lycopersicon esculentum (tomato). Sequence analysis reveals a high degree of similarity between the TUB13 cDNA, and a human S-adenosylmethionine decarboxylase gene. The predicted TUB8 peptide sequence shows several repeats of alanine, glutamate and proline which suggests a structural role for the encoded protein.

Adenosylmethionine Decarboxylase↗
PubMed1992

Ty1-copia group retrotransposons and the evolution of retroelements in the eukaryotes.

Ty1-copia group retrotransposons are among the best studied transposable elements in the eukaryotes. This review discusses the extent of these transposons in the eukaryote kingdoms and compares models for the evolution of these genetic elements in the light of recent phylogenetic data. These data show that the Ty1-copia group is widespread among invertebrate eukaryotes, especially in the higher plant kingdom, where these genetic elements are unusually common and heterogeneous in their sequence. The phylogenetic data also suggest that the present day spectrum of Ty1-copia group retrotransposons has been influenced both by divergence during vertical transmission down evolving lineages and by horizontal transmission between distantly related species. Lastly, the factors affecting Ty1-copia group retrotransposon copy number and sequence heterogeneity in eukaryotic genomes and the effects of transpositional quiescence and defective retrotransposons upon evolution of Ty1-copia group retrotransposons are discussed.

Amino Acid Sequence↗
PubMed1992

Extreme heterogeneity of Ty1-copia group retrotransposons in plants.

We have used the polymerase chain reaction to analyse Ty1-copia group retrotransposons of flowering plants. All eight species studied contain reverse transcriptase fragments from Ty1-copia group retrotransposons. Sequence analysis of 31 subcloned fragments from potato reveals that each is different from the others, with predicted amino acid diversities between individual fragments varying between 5% and 75%. Such sequence heterogeneity within a single species contrasts strongly with the limited diversity seen in such retrotransposons in yeast and Drosophila. The fragments from the other seven plant species examined are also heterogeneous, both within and between species, showing that this is a general property of this transposon group in plants. Phylogenetic analysis of all these sequences reveals that many of them fall into subgroups which span species boundaries, such that the closest homologue of one sequence is often from a different species. We suggest that both vertical transmission of Ty1-copia group retrotransposons within plant lineages and horizontal transmission between different species have played roles in the evolution of Ty1-copia group retrotransposons in flowering plants.

Amino Acid Sequence↗
PubMed1992

A Ty1-copia group retrotransposon sequence in a vertebrate.

We have used the polymerase chain reaction (PCR) to isolate a sequence characteristic of a Ty1-copia group retrotransposon from the genome of the herring (Clupea harengus). This is the first Ty1-copia group retrotransposon sequence described in a vertebrate. Phylogenetic comparison of this sequence with other members of this group of retrotransposons shows that it resembles more closely some Ty1-copia group members from Drosophila melanogaster than other group members in plants and fungi. These observations provide further evidence that the Ty1-copia group LTR retrotransposons span many of the major eukaryote species boundaries, suggesting that horizontal transmission between different species has played a role in the evolution of this retrotransposon group.

Amino Acid Sequence↗
PubMed1991

The retrotransposon copia regulates Drosophila gene expression both positively and negatively.

The D. melanogaster transposable element copia is structurally similar to retroviral proviruses. We have asked whether copia encodes regulatory functions which have been observed in certain other proviruses. We have introduced reporter constructs based on the copia promoter and other Drosophila promoters into Drosophila cells and asked if copia has any affect on their expression. We find that, whilst copia negatively regulates expression from its own promoter, it also positively regulates expression from the larval serum protein 1 promoter. Analysis of RNA suggests that both regulatory functions occur by post-transcriptional mechanisms.

Animals↗
PubMed1990

The retrotransposon copia controls the relative levels of its gene products post-transcriptionally by differential expression from its two major mRNAs.

All retroviruses and retrotransposons studied to date regulate the relative levels of gag and pol/int gene products post-transcriptionally from a single mRNA. In these genetic elements the production of protein encoded by the pol and int genes is attenuated by a translational stop or frameshift in the reading frame preceding their coding regions in the mRNA. We show here that the Drosophila retrotransposon copia also produces lower amounts of gene products from its int/pol region than gag region but this is achieved by a mechanism which is novel for this class of genetic element. We show by the use of gene fusion constructs that the subgenomic 2 kilobase copia RNA, encoding gag products, is expressed as protein in cultured cells at least ten-fold more efficiently than the full genome length RNA, which additionally contains the pol and int open reading frames.

Animals↗
PubMed1990

Complete reversions of a gypsy retrotransposon-induced cut locus mutation in Drosophila melanogaster involving jockey transposon insertions and flanking gypsy sequence deletions.

We have analysed the structures of three phenotypic revertant alleles of a gypsy retrotransposon-induced mutation at the cut locus of Drosophila melanogaster. All three revertants are associated with the insertion of jockey transposons into a common region of gypsy. Two of these alleles are complete reversions to wild type. One complete revertant (ct+D) is derived from a third allele, a partial revertant (ctMRpD) by a deletion of part of the gypsy sequence flanking the jockey transposon. Sequence differences between the jockey elements in ctMRpD and ct+D suggest that this deletion may have been created by the insertion of a second jockey near to the first, followed by recombinational excision of a composite jockey and the region between the two genetic elements. The other complete revertant also carries a deletion of gypsy DNA flanking the jockey insertion. The deleted regions of both complete revertants and the target region for all the jockey insertions contain a repeated sequence that resembles a transcriptional enhancer. The strength of the cut phenotype in these mutants correlates with the proportion of this region remaining near the gypsy transcriptional start site, suggesting that the jockey insertions relieve the gypsy-induced mutation at cut by interfering with a region which is required for the transcriptional competence of gypsy.

Alleles↗
PubMed1989

The transcriptional control regions of the copia retrotransposon.

We have analysed the sequence elements that control expression of the copia retrotransposon. Expression of copia fusion constructs containing DNA sequence deletions and rearrangements was assayed by transient expression analysis. Progressive deletion and linker substitution identifies two regions on either side of the major transcriptional start sites in the copia long terminal repeat These regions are both required for high level expression in a cultured Drosophila melanogaster cell line but only the upstream region is required for copia expression in a Drosophila hydei cell line. A third control region lies downstream of the long terminal repeat in a region previously believed to contain no cis-acting regulatory sequences. We show by displacement and inversion of this region that it contains a transcriptional enhancer.

Animals↗
PubMed1989

Instability in the ctMR2 strain of Drosophila melanogaster: role of P element functions and structure of revertants.

Simultaneous multiple transpositions and long-term genetic instability have been described in the ctMR2 strain of Drosophila melanogaster and its derivatives. This strain originated from a cross that was dysgenic in the P-M system. While spontaneous instability declined over 2 years, instability has been reactivated by backcross to the progenitor P element bearing strain MRh12/Cy. We show here using germline transformation that active P factor alone cannot mimic the effect of this cross, suggesting that MRh12/Cy contains some other activator. In addition, we have observed that ct+ exceptional progeny arise in the F1 as well as the F2 generations. Molecular analysis of X chromosomes from some ct+ progeny indicates that phenotypic reversion of the ct mutation can arise through two unrelated mechanisms.

Alleles↗
PubMed1987

A novel GC-rich dispersed repeat sequence in Drosophila melanogaster.

A novel family of dispersed repeat sequences from Drosophila melanogaster is described. Sequence analysis of two members of this family show them to contain greater than 75% GC bases. These are comprised of multiple repeats of GGX triplets interspersed occasionally with CGPy and TTPy. Southern blotting shows that these repeats are not transposable elements. Twenty four homologous recombinants have been localised by in situ hybridization to seven sites in the Drosophila genome. Polyadenylated RNAs homologous to this repeat family are expressed in a complex pattern which is developmentally regulated. We suggest that this family encodes a set of glycine-rich domains in Drosophila proteins.

Animals↗
PubMed1986

The termini of extrachromosomal linear copia elements.

Extrachromosomal linear copia elements were isolated and cloned from cultured Drosophila melanogaster cells. Four full length copia recombinants were characterised. Each contains a full sized copy of copia bounded by its direct repeats. The sequences of the ends of the copia inserts were determined. All of the termini are identical to those of genomic copias except for a single base deletion from one end of one clone. These results suggest that the priming for initiation of copia reverse transcription differs from normal retroviral models. Southern blot analysis of uncloned full length copia circles shows that approximately 50% correspond to the predicted circularisation product of such linears. We propose that this class of circles and the linear DNAs described here are precursors to integrated genomic copia elements.

Animals↗
PubMed1983

The origin of extrachromosomal circular copia elements.

Cloned extrachromosomal circular copia elements were studied by nucleotide sequence and restriction enzyme analysis to determine their mechanisms of formation and their possible roles in copia transposition. Rearranged circular copias containing inverted segments flanked by 5 bp sequence duplication were observed, suggesting that circular copias are capable of integrating into their own sequences. Such copia circles are analogous to similarly rearranged retrovirus circles, strengthening the relationship between the copia-like elements and retroviruses. There is marked sequence heterogeneity at the junction between the fused terminal direct repeats of seven copia circles. The junctions contain 0-15 bp inserts, the sequences of which are inconsistent with the creation of these particular molecules by reverse transcription.

Animals↗
PubMed1981

The 5' termini of RNAs encoded by the transposable element copia.

The 5' termini of copia-specific RNAs in Drosophila melanogaster tissue culture cells were determined by S1 nuclease mapping and cap analysis. Both major copia RNAs share an identical set of heterogeneous 5' ends. Three major cap 1 structures M7GpppCmpUp, M7GpppCmpCp and M7GpppGmpUp together with several other minor caps were found. Almost all the 5' termini, as judged by S-1 nuclease mapping, were located either in a pyrimidine-rich part of the terminal direct repeat or apparently outside of the copia element, suggesting that a proportion of copia transcripts derive from promoters external to the genetic element.

Animals↗