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At least 973 records · Page 54Linked to original sources
PubMed1997

A trans-acting modifier causing extensive overexpression of genes in Drosophila melanogaster.

A modified P element-induced mutation at cytological position 47A11-12 on chromosome 2 of Drosophila melanogaster inversely alters the expression of several genes examined. In addition, the effect is nearly exponential in some developmental stages for five of the eleven genes tested (white, brown, scarlet, rudimentary and vermilion). Interestingly, the exponential overexpression of the transcripts leads to a sexually dimorphic effect, with female levels being greater. The white allele specificity analysis and the lack of response of an Alcohol dehydrogenase promoter-white reporter fusion construct suggest that the 5' regulatory sequences of white are required for the response. Because the cases of extreme overexpression are more prominent in females than males, the locus is termed Ultra female overexpression (Ufo). Ufo revertants have no discernible effect on the transcript levels, indicating that the mutation is responsible for the modulation of the tested genes. Regulatory genes such as Ufo may contribute to the wide fluctuations in gene expression between tissues and stages of development.

ATP-Binding Cassette Transporters↗
PubMed1997

Germ line transposition of the copia retrotransposon in Drosophila melanogaster is restricted to males by tissue-specific control of copia RNA levels.

Germ line transposition rates of the retrotransposon copia were directly measured in males and females of an inbred Drosophila melanogaster line, 2b3, which is highly polymorphic for copia insertion sites. The elevated germ line transposition rate of copia in this line (3-8 x 10(-3) per generation per element) is confined to males, with transposition in females being undetectable under the conditions of the experiment but at most 50-fold lower than the rate for males. To determine the molecular basis of this effect, copia RNA levels were measured in whole bodies and germ lines of male and female flies of both the unstable 2b3 line and a stable line, Oregon RC-iso, which shows normal rates of copia transposition. Both male and female 2b3 flies contain much more copia RNA than flies of the stable line. However, 2b3 male germinal tissues contain much higher levels of copia RNA than the equivalent female tissues. The highest copia expression is detected in maturing primary spermatocytes. Our data show that high rates of germ line copia transposition are restricted to males by tissue-specific control of RNA levels and suggest that transposition of copia only occurs in fly tissues containing more than a relatively high threshold level of copia RNA.

Animals↗
PubMed1998

The RNA product of the Doc retrotransposon is localized on the Drosophila oocyte cytoskeleton.

The Doc retrotransposon of Drosophila melanogaster encodes an RNA with two open reading frames (ORFs). ORF1 encodes an RNA-binding protein and overlaps with ORF2, which encodes a reverse transcriptase. Retrotransposons transpose to new positions in the genome via a full-length RNA intermediate. Because they do not encode a coat protein, transposition is limited to single cell lineages. In this paper we present evidence that shows that the highest levels of Doc transcripts are found in the adult male and female germ-line. The Doc RNA is localized in the region of the cytoskeleton of the Drosophila oocyte. The biological significance of this is unknown, but we discuss some of the possibilities.

Animals↗
PubMed1998

Role of multiple trans-acting regulators in modifying the effect of the retrotransposon copia on host gene expression in Drosophila.

We have examined the combinatorial effect of five trans-acting modifiers of white apricot, a copia retrotransposon-induced allele of the white eye-color gene. Quantitation of steady-state levels of copia transcripts for various modifier combinations at different developmental stages using northern analysis reveals nonadditivity and epistasis. Interestingly, extensive overexpression of copia transcripts caused by the modifier Ufo is altered up to twofold in level when Ufo is combined with either of the modifiers Mow or Wow. In general, double combinations do not elevate the level of copia transcripts more than twofold. The role of suppressor mutations in modifying the effect of transposable elements on host gene expression is discussed.

Animals↗
PubMed1998

Structure of the Chlorella Zepp retrotransposon: nested Zepp clusters in the genome.

Zepp elements found in the telomeric region of Chlorella chromosomes show the characteristic features of non-viral (LINE-like) retrotransposons, including a poly(A) tail, 5' truncations, a retroviral reverse transcriptase-like ORF and flanking target duplications. We have isolated and characterized a full-length Zepp element (8943 bp long) from Chlorella chromosome V. Some peculiar features of this element, including nested integration, two ORF structures, a long 3' noncoding region and a possible promoter region are compared with those of the Drosophila telomeric retrotransposons HeT-A and TART. The Chlorella chromosome-Zepp system appears to represent an intermediate stage between canonical telomerase-telomeres and Drosophila retrotransposon-telomeres.

Amino Acid Sequence↗
PubMed1998

Molecular characterization of a retrotransposon in Drosophila melanogaster, nomad, and its relationship to other retrovirus-like mobile elements.

The nomad element was identified as a retrovirus-like transposon in Drosophila melanogaster. DNA sequence analysis showed that the nomad element contains three long ORFs that are similar to the gag, pol and env genes of retrovirus- and the copia-like elements of D. melanogaster. The nomad element terminates with 519-bp terminal repeats, each of which contains eukaryotic consensus transcription initiation and termination signals. nomad elements are located at approximately 10-15 sites within the euchromatic arms of the genome and at the chromocenter, as shown by in situ hybridization. The host DNA sequence TANA was found to be duplicated on each side of the nomad element and appears to be a preferential target site for insertion of nomad elements. Analysis of the zinc finger motif in the pol gene product of retrotransposons known to have target site preference suggests involvement of the integrase subunit in target site selection for those retrotransposons that display insert site specificity. A comparison of the predicted amino acid sequence of the pol-like genes of several known retrotransposons was made and the phylogenetic relationship between nomad and other retrovirus-like mobile elements was determined. It was clear from this conceptual protein analysis and from analysis of their structural characteristics that retrotransposons of the gypsy class can be generally classified as members of one of two distinct groups. The phylogenetic relationships of these groups are also discussed.

Amino Acid Sequence↗
PubMed1999

Identification and characterization of novel retrotransposons of the gypsy type in rice.

We found that two DNA fragments, which were obtained from Oryza sativa L cv. IR36 by PCR using degenerate primers designed for amplification of a rice gene, showed homology with the rt gene encoding reverse transcriptase of the Drosophila retrotransposon gypsy. We named the element from which they originated RIRE3 (for rice retrotransposon No. 3) and analyzed it further by isolating various clones containing segments of RIRE3. Nucleotide sequencing of the clones revealed that RIRE3 has LTRs (2316 bp) and that the internal sequence (5775 bp) includes a large ORF with gag and pol regions; the pol region includes the rt gene as well as the int gene encoding integrase in this order, as in gypsy. Interestingly, the region upstream of gag in RIRE3 contained another open reading frame, here called orf0, which does not exist in gypsy or in other retrotransposons related to it. In the course of characterizing RIRE3, we obtained a further clone, which showed less homology with the pol region of RIRE3. This clone was found to be derived from another gypsy-type retrotransposon (named RIRE8) containing the LTR sequence and orf0 both of which were only weakly homologous to that in RIRE3. Further characterization of RIRE8 revealed that there were actually two subtypes of RIRE8 (named RIRE8A and RIRE8B), which show little homology to each other in the orf0 region. Although the LTRs of RIRE3 and RIRE8 elements show very weak homology with each other, there exists a conserved sequence at their termini. We therefore carried out PCR using primers which hybridize to the rt gene of RIRE3, and total genomic DNA from various monocot and dicot plants as templates, and found that a family of RIRE3 elements was present in all plants tested.

Amino Acid Sequence↗
PubMed1999

Non-LTR retrotransposons (LINEs) as ubiquitous components of plant genomes.

During the course of work aimed at isolating a rice gene from Oryza australiensis by PCR, the oligonucleotide primers used were found to generate a fragment that showed sequence homology to the endonuclease (EN) region of the maize non-LTR retrotransposon (LINE) Cin4. We carried out further PCRs using oligonucleotide primers that hybridized to these sequences, and found that they amplified several fragments, each with homology to the EN regions, from Oryza sativa cv. Nipponbare as well as O. australiensis. We mapped the approximate locations of two rice LINE homologues by screening clones in a YAC library made from a rice (O. sativa) genome, and found that each homologue was present in a low copy number apparently at nonspecific regions on rice chromosomes. We then carried out PCR using degenerate oligonucleotide primers which hybridized to the rice LINE homologues and Cin4 to ascertain whether LINE homologues are present in a variety of members of the plant kingdom, including angiosperms, gymnosperms, bracken, horsetail and liverwort. Cloning and nucleotide sequencing revealed that 53 clones obtained from 27 out of 33 plant species contained LINE homologues. In addition to these homologues, we identified four homologues with EN regions in the Arabidopsis thaliana genome by a computer search of databases. The nucleotide sequences of almost all the LINE homologues were greatly diverged, but the derived amino acid sequences were well conserved, and all contained glutamic acid and tyrosine residues at almost the same relative positions as in the the active site regions of AP (apurinic/apyrimidinic)-endonucleases. The EN regions in the LINE homologues from closely related plant species show a closer phylogenetic relationship, indicating that sequence divergence during vertical transmission has been a major influence upon the evolution of plant LINEs.

Amino Acid Sequence↗
PubMed1999

A novel simple satellite DNA is colocalized with the Stalker retrotransposon in Drosophila melanogaster heterochromatin.

In the T(1:2)dor(var7) multibreak rearrangement the distal 1A-2B segment of the X chromosome of Drosophila melanogaster is juxtaposed to an inverted portion of the heterochromatin of chromosome 2. Analysis of mitotic chromosomes by a series of banding techniques has permitted us precisely to locate the heterochromatic breakpoint of this translocation in the h42 region of 2R. Cloning and sequencing of the eu-heterochromatic junction revealed that the translocated 1A-2B fragment is joined to (AACAC)n repeats, which represent a previously undescribed satellite DNA in D. melanogaster. These repeated sequences have been estimated to account for about 1 Mb of the D. melanogaster genome. The repeats are located mainly in the Y chromosome and in the heterochromatin of the right arm of chromosome 2 (2Rh), where they are colocalized with the Stalker retrotransposon.

Animals↗
PubMed1999

Degenerate MAGGY elements in a subgroup of Pyricularia grisea: a possible example of successful capture of a genetic invader by a fungal genome.

The LTR-retrotransposon MAGGY is found sporadically in isolates of Pyricularia grisea (Magnaporthe grisea). Based on a dendrogram constructed by RFLP analysis of rDNA, isolates that carry MAGGY elements were classified into a single cluster that comprised four rDNA types. However, in a few members of this cluster, exemplified by isolates from common millet (Panicum miliaceum), the MAGGY element has distinct features. Southern analysis suggested that these isolates possessed a single copy of a MAGGY-related sequence whose restriction map differed from that of MAGGY itself. Sequence analysis revealed that the MAGGY-related sequence was a degenerate form of MAGGY, characterized by numerous C:G to T:A transitions, which have often been reported to result from RIP (Repeat-induced point mutation) or RIP-like processes. However, the favored target site for C:G to T:A transitions in this fungus, determined by examining a total of 501 sites, was (A/T)pCp(A/T), which differs from that for the RIP process originally reported in Neurospora (CpA), and from that reported in Aspergillus (CpG). The fact that certain members of the cluster of MAGGY carriers retain a single copy of a degenerate MAGGY element implies that the ancestor of these isolates successfully "captured" the invading MAGGY element.

Amino Acid Sequence↗
PubMed1999

HeT-A telomere-specific retrotransposons in the centric heterochromatin of Drosophila melanogaster chromosome 3.

We have isolated two yeast artificial chromosome (YAC) clones from Drosophila melanogaster that contain a small amount of dodeca satellite (a satellite DNA located in the centromeric region of chromosome 3) and sequences homologous to the telomeric retro-transposon HeT-A. Using these YACs as probes for fluorescence in situ hybridization to mitotic chromosomes, we have localized these HeT-A elements to the centric heterochromatin of chromosome 3, at region h55. The possible origin of these telomeric elements in a centromeric position is discussed.

Animals↗
PubMed1999

Heterogeneity of the internal structure of PDR1, a family of Ty1/copia-like retrotransposons in pea.

We characterised the extent of heterogeneity among PDR1 elements, a Ty1/copia-like retrotransposon family in pea, by restriction mapping and PCR with primers designed to amplify four functional domains. The data suggest that two main subfamilies of PDR1 differ in the size of their 5'-region. There are also sequence variants and rearranged copies which include a wide range of deletions of different sizes and deletions combined with insertions of host DNA, or inversions of various regions of the retrotransposon. A deletion hotspot has been found at nucleotide position 394, where buffer sequences of 26 bp and 38 bp containing microsatellite motifs have been generated. There is more heterogeneity in the gag domain of PDR1 than in other functional domains, and the extent and pattern of this diversity was assessed among 56 Pisum accessions. We found a higher rate of rearrangement and sequence variation within the gag domain of PDR1 in P. fulvum and P. abyssinicum accessions than would be expected from the degree of insertion site polymorphism. A neighbour-joining phylogenetic tree constructed for gag sequences has a similar branching pattern to the equivalent insertion site tree, implying that the PDR1 family and its gag domain have coevolved with the pea genome. Combining both trees revealed clear and distinct subgroups among the Pisum ssp.

Base Sequence↗
PubMed2001

DNA methylation inhibits expression and transposition of the Neurospora Tad retrotransposon.

Tad is a LINE-like retrotransposon of the filamentous fungus Neurospora crassa. We have analyzed both expression and transposition of this element using strains with a single copy of Tad located in the 5' noncoding sequences of the am (glutamate dehydrogenase) gene. Tad in this position has been shown to carry a de novo cytosine methylation signal which causes reversible methylation of both Tad and am upstream sequences. Here we find that methylation of the Tad sequences inhibits both Tad expression and transposition. This inhibition can be relieved by the use of 5-azacytidine, a drug which reduces cytosine methylation, or by placing the Tad/am sequences in a dim-2 genetic background.

Azacitidine↗
PubMed2001

A family of DNA repeats in Aspergillus nidulans has assimilated degenerated retrotransposons.

In the course of a chromosomal walk towards the centromere of chromosome IV of Aspergillus nidulans, several cross-hybridizing genomic cosmid clones were isolated. Restriction mapping of two such clones revealed that their restriction patterns were similar in a region of at least 15 kb, indicating the presence of a large repeat. The nature of the repeat was further investigated by sequencing and Southern analysis. The study revealed a family of long dispersed repeats with a high degree of sequence similarity. The number and location of the repeats vary between wild isolates. Two copies of the repeat contained degenerated long-terminal-repeat (LTR) retrotransposons, which were named Dane1 and Dane2 (for Degenerated Aspergillus nidulans element). The pattern of degeneration suggested that a process similar to the repeat-induced point-mutation (RIP) phenomenon, first described in Neurospora crassa, may have operated in A. nidulans. The data indicate that this family of repeats has assimilated mobile elements that subsequently degenerated but then underwent further duplications as a part of the host repeats.

Amino Acid Sequence↗
PubMed2001

Retrolyc1 subfamilies defined by different U3 LTR regulatory regions in the Lycopersicon genus.

Retrolycl, a Ty1/copia-like element, was originally isolated from the Lycopersicon peruvianum genome and shown to be present also in other Lycopersicon species. It shares extensive similarities with Tntl, except in its U3 regulatory region. In order to evaluate Retrolycl diversity, we analyzed partial sequences including both coding domains and the U3 regulatory region in four different species of the Lycopersicon genus. Two Retrolycl subfamilies defined by different U3 regions were identified. RetrolyclA is most abundant in L. peruvianum and L. hirsutum, while Retrolyc1B is distributed in all four species studied here. The RetrolyclA U3 region contains tandemly repeated elements of 53 bp. Transient expression analysis suggests that Retrolyc1A is a transcriptionally active family, and that the repeated motifs found in its U3 region are important transcriptional regulatory elements.

Base Sequence↗
PubMed2001

Heat shock, copper sulfate and oxidative stress activate the retrotransposon MAGGY resident in the plant pathogenic fungus Magnaporthe grisea.

MAGGY is a gypsy-like retrotransposon isolated from the plant pathogenic fungus Magnaporthe grisea. The ability of various stresses to activate MAGGY was tested in the original and in a heterologous host (Colletotrichum lagenarium), using beta-glucuronidase (GUS) as a reporter. The MAGGY promoter was activated in M. grisea by either heat shock, copper sulfate, or oxidative stress, but not by the antifungal substance p-coumaric acid. Transcriptional up-regulation of MAGGY RNA was also observed following heat shock and oxidative stress. The MAGGY promoter remained responsive to the above-mentioned stresses when transformed into a M. grisea isolate that had no endogenous MAGGY elements. In C. lagenarium, however, the MAGGY promoter showed only basal expression of GUS and no further up-regulation was induced by any of the stress treatments, suggesting that the stress-responding cis-element(s) in the MAGGY promoter is not functional in a wider range of fungi. The relationship between the activation of MAGGY by stress and phenotypic diversification in M. grisea, including variations in pathogenicity, is discussed.

Amino Acid Sequence↗
PubMed2000

Age and origin of the FCMD 3'-untranslated-region retrotransposal insertion mutation causing Fukuyama-type congenital muscular dystrophy in the Japanese population.

Fukuyama-type congenital muscular dystrophy (FCMD), an autosomal recessive disorder with a high prevalence in the Japanese population, is characterised by severe muscular dystrophy associated with brain malformation (cortical dysgenesis) and mental retardation. In Japan, 87% of FCMD-bearing chromosomes carry a 3-kb retrotransposal insertion of tandemly repeated sequences within the disease gene recently identified on chromosome 9q31, and most of them share a common founder haplotype. FCMD is the first human disease known to be caused primarily by an ancient retrotransposal integration. By applying two methods for the study of linkage disequilibrium between flanking polymorphic markers and the disease locus, and of its decay over time, the age of the insertion mutation causing FCMD in Japanese patients is calculated to be approximately 102 generations (95% confidence interval: 86-117 g), or slightly less. The estimated age dates the most recent common ancestor of the mutation-bearing chromosomes back to the time (or a few centuries before) the Yayoi people started migrating to Japan from the Korean peninsula. This finding makes the molecular population genetics of FCMD understandable in the context of Japan's history and the founder effect consistent with the prevalent theory on the origins of the modern Japanese population.

Asian People↗
PubMed1997

Interethnic polymorphism of EWS intron 6: genome plasticity mediated by Alu retroposition and recombination.

The EWS gene has been identified as being systematically translocated in Ewing's sarcoma. In order to ascertain the basis of a marked interethnic difference in the incidence of Ewing's sarcoma, intron 6 of EWS, which is located near the translocation breakpoint region (EWSR1), was characterized. Sequence analysis of the entire intron 6 region revealed a very high density of Alu elements. Most of these Alu sequences could be classified in previously described subfamilies, facilitating delineation of an evolutionary model that involves successive retroposition events. According to this model, the EWS intron 6 region progressively expanded until about 5 million years ago. More recently (10(5) years ago), in part of the human population, the size of this region decreased by over 50% as the result of a homeologous recombination between two Alu sequences, which removed 2480 bp. This rare allele has only been observed in individuals of African origin, a population that is characterized by the lowest incidence of Ewing's sarcoma.

Alleles↗