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At least 325 records · Page 18Linked to original sources
PubMed1996

[Molecular cloning of mutant loci obtained by microinjection of oncogenic viral DNA into polar plasma of early embryos of Drosophila melanogaster].

Mutations WRSV-1 and WRSV-4 obtained by microinjection of RSV cDNA into early embryos of Drosophila melanogaster were shown to be caused by the insertion of mobile elements jockey and B104(roo), respectively partial reversion of WRSV-lr arose due to the deletion of a 5'-region of mobile element jockey and of the adjacent region of the genomic DNA. It is concluded that genetic instability induced by oncoviral DNAs is based on the ability of these DNAs to induce transpositions of various mobile elements in the recipient genome.

Animals↗
PubMed2003

Genomic structure of the NtPDR1 gene, harboring the two miniature inverted-repeat transposable elements, NtToya1 and NtStowaway101.

Here we report the genomic structure including the promoter sequence and coding region of NtPDR1 (Nicotiana tabacum Pleiotropic Drug Resistance 1), which is an elicitor-responsive gene encoding an ATP binding cassette (ABC) transporter that might be involved in the defense response in tobacco, as we reported recently. The NtPDR1 gene consists of 20 exons and 19 introns. Among the introns, the first and fifth are much larger than the others and harbor typical miniature inverted-repeat transposable elements (MITEs). One of the MITE elements in the first intron, termed NtToya1, belongs to the Toya family that was recently described in rice, while the other element in the fifth intron, termed NtStowaway101, shows high homology with the Stowaway elements of the IS630-Tc1-mariner family. Many of the genes we found to harbor Toya and Stowaway elements in Nicotiana species by BLAST search are also involved in stress responses or plant-pathogen interactions. The existence of putative cis-elements (a GCC box, three W boxes, and several JA-responsive elements) in the promoter region supports our previous finding that this gene is strongly inducible by elicitation and methyljasmonate, and that this ABC transporter might be essential for plant defense responses. Furthermore, Southern blot analysis and PCR amplification of the introns harboring the MITE-like elements from genomic DNA of three Nicotiana species suggests that NtPDR1 originated from N. sylvestris.

Base Sequence↗
PubMed1998

Analysis of flower pigmentation mutants generated by random transposon mutagenesis in Petunia hybrida.

Fifty new flower pigmentation mutants in Petunia hybrida using endogenous transposable elements (TEs) as a mutagen were generated. Forty-six mutants displayed somatic and sporogenic instability indicating that they were caused by a TE. Phenotypic analysis showed that the mutation altered either anthocyanin biosynthesis (40 alleles for seven loci), the intracellular pH of petals (six alleles for three loci) or the shape of petal cells (two alleles for two loci). To identify the TEs responsible for the mutations, the authors subjected 16 alleles of the anthocyanin-3 (an3) locus, encoding flavanone 3 beta-hydroxylase, to molecular analysis. This showed that 11 out of 12 unstable an3 alleles harboured TE insertions of a single family, dTph1, while one allele harboured a new 177 bp TE designated dTph2. In addition, the authors found one an3 allele (an3-W138A) in which a dTph1 element had inserted 30 bp upstream the translation start, without inactivating the gene. This 'cryptic' element was responsible for the creation of a stable recessive (untagged) an3 allele, where a large rearrangement inactivated the gene. These findings indicate that mutants for novel loci are most likely tagged by dTph1 elements opening the way for their isolation.

Alleles↗
PubMed2004

Characterization of an unusual Ds transposable element in Arabidopsis thaliana: insertion of an abortive circular transposition intermediate.

The maize Ac/Ds transposable elements, which belong to the hAT transposon superfamily, are widely used as insertional mutagens in numerous plant species. Molecular studies suggest that Ac/Ds elements transpose in a conservative non-replicative fashion; however the molecular mechanism of transposition remains unclear. We describe here the identification of an unusual Ds element, Ds-mmd1 , in a transgenic Arabidopsis line. Ds-mmd1 is rearranged relative to the original Ds element, such that the original 5' and 3' ends are internal and previously internal sequences are the new 5' and 3' termini of Ds-mmd1 . Short duplications of plant genomic DNA and Ds sequences are present at the Ds-mmd1 junctions, suggesting that a circular Ds molecule was part of the events that created the Ds-mmd1 element. In addition, a revertant analysis on mmd1 plants demonstrated that Ds-mmd1 can be eliminated from the genome in an Ac -dependent process.

Arabidopsis↗
PubMedDate not supplied

The transposable element IS4712 prevents S-layer gene (sbsA) expression in Bacillus stearothermophilus and also affects the synthesis of altered surface layer proteins.

Cell surface (S)-layer protein synthesis in Bacillus stearothermophilus PV72/p6 is blocked when cells are grown at elevated temperature. From a culture exhibiting the S-layer-negative phenotype, the S-layer deficient mutant T5 (SbsA-) was isolated. Genetic analysis of the S-layer-encoding gene (sbsA) of mutant T5 revealed an insertion element (IS4712) integrated into the upstream regulatory region of the S-layer gene, thereby blocking sbsA transcription. The insertion element consists of 1371 base pairs which are flanked by two perfect inverted terminal repeats. Sequence similarity to other transposases of the IS4 family was detected. DNA-DNA hybridizations demonstrated that multiple homologues of IS4712 were also present within the genomes of several other thermophilic bacillus isolates. Attempts to isolate SbsA+ revertants failed. Instead, cells with altered surface proteins were detected. The synthesis of the altered S-layer proteins was correlated with the presence of IS4712 along with the occurrence of deletions in the sbsA coding region. Furthermore imprecise excision of IS4712 was detected. This work demonstrated that B. stearothermophilus is able to express at least four different S-layer proteins and that blocking of sbsA transcription by the insertion element IS4712 is associated with the expression of altered surface proteins.

5' Untranslated Regions↗
PubMed1986

The anterobithorax and bithorax mutations of the bithorax complex.

The anterobithorax (abx) and bithorax (bx) genes together direct the development of the posterior second and anterior third thoracic segments of the fruit fly. We have characterized the phenotypes and DNA lesions of 19 abx and bx alleles. abx and bx mutations differ both in the nature and location of their DNA lesions, forming two clusters within a relatively small region of the Ultrabithorax transcription unit. Correlation between phenotype and DNA lesion suggests the presence of two or more genetic elements in this region distinct from the Ultrabithorax transcript. Mutant transformations do not strictly obey segmental or parasegmental boundaries. Most of the bx mutations result from insertions of the mobile element gypsy. The strength of these alleles varies in a regular way dependent on the position and orientation of the gypsy element. We propose models for gypsy element action and bithorax complex expression in the light of these results.

Animals↗
PubMed2000

The novel insertion sequences IS1417, IS1418, and IS1419 from Burkholderia glumae and their strain distribution.

Three insertion sequences, IS1417, IS1418, and IS1419, were isolated from Burkholderia glumae (formerly Pseudomonas glumae), a gram-negative rice pathogenic bacterium, on the basis of their abilities to activate the expression of the neo gene of the entrap vector pSHI1063. The 1335-bp IS1417 element with 17-bp imperfect terminal inverted repeats was found to be flanked by 5-bp direct repeats of the vector sequence. IS1418 is 865 bp in length and carries 15-bp inverted repeats with a target duplication of 3 bp. The 1215-bp IS1419 sequence is bounded by the 36-bp terminal inverted repeats of the element and 7-bp direct repeats of the vector sequence. IS1417 and IS1418 belong to the IS2 subgroup of the IS3 family and the IS427 subgroup of the IS5 family, respectively, whereas IS1419 does not appear to be a member of any known IS family. Southern blot analysis of DNAs from B. glumae field isolates indicated that those IS elements are widely distributed, but the host range of the three IS elements appears to be limited to B. glumae and some other related species such as B. plantarii. The polymorphisms exhibited in B. glumae isolates suggest that those elements are useful for molecular epidemiological studies of B. glumae infections.

Amino Acid Sequence↗
PubMed2001

The wild-type conformation of the Mos-1 inverted terminal repeats is suboptimal for transposition in bacteria.

The two inverted terminal repeats (ITRs) flanking the Mos-1 mariner element differ in sequence at four positions. Gel retardation experiments indicated that each of these differences has a significant impact on the quality of the interaction between the ITR and the Mos-1 transposase. We showed that the transposase binds to the 3' ITR better than to the 5' ITR. The results of transposition assays performed in Escherichia coli indicated that these differences have an influence on the rate of transposition and the stability of the transposition products. Finally, we find that the wild-type configuration of the Mos-1 element, with one 5' ITR and one 3' ITR, is less efficient for transposition in bacteria than that of an element having two 3' ITRs.

Base Sequence↗
PubMed1979

Inceptor and origin of DNA replication in lambdoid coliphages. I. The lambda DNA minimal replication system.

In a pBR313-lambda dv hybrid plasmid system, stepwise deletion and serial cloning procedures have led to a functional dissection of the DNA replication region of lambdoid bacteriophages lambda, 434 and 21. A simple system for initiation of DNA replication has been detected within lambdoid replicator DNAs, which is active in the absence of several normal replication elements, including the origin of replication (ori) and product of gene O. This "minimal" (or "mini") initiation system depends on the p0 or substitute leftward promoter in conjunction with the newly discovered "inceptor" (ice) element, which is located within the cII gene. Even the fragments containing ori are unable to initiate replication in these hybrid plasmids as long as fragments containing ice are missing. The base sequence of ice resembles transcriptional terminators and it appears to control both termination of primer RNA and inception of daughter strand DNA synthesis. Initiation in the p0-ice mini system of lambda or 21 phages requires the gene P product. Hwever, mini replication of 434 DNA hybrid plasmids required neither O nor P proteins, although there are only two single-base changes in the 434 inceptor sequence. The mini system is repressed by the elements of the maximal lambda replication system, as described in the accompanying publication.

Bacteriophage lambda↗
PubMed2004

Mobile DNA elements in the gas vesicle gene cluster of the planktonic cyanobacteria Microcystis aeruginosa.

Insertion sequences (IS) have been characterized in Microcystis aeruginosa gas vesicle-deficient mutants. ISMae4, a homolog of the cyanobacterial IS702, belongs to the IS5 family, subgroup ISL2. ISMae2 and ISMae3 display typical IS features and express a transposase of the IS4 and IS1 family, respectively. ISMae1 exhibits a more complex genetic structure and harbours a degenerated transposase gene distantly related to IS1 elements. Hybridizations with IS-specific DNA probes suggest that transposition of ISMae2 and ISMae3 occurred by a replicative-type mechanism. To our knowledge this is the first report showing that IS1 elements can be mobile in cyanobacteria.

Amino Acid Sequence↗
PubMedDate not supplied

Estimation of the rate of cloned gene integration via retrotransposition.

A theoretical method has been developed to estimate retrotransposition (integration) rates of the Saccharomyces cerevisiae Ty3 and Ty1 retrotransposons bearing heterologous and homologous genes (neor, HIS3). The method is based on population growth modeling and Lea and Coulson's maximal likelihood method for mutation rate estimation (Lea and Coulson, 1949). This method has allowed us to examine directly retrotransposition rates of GAL-regulated marked Ty3 and Ty1 elements into the whole yeast genome, not just a particular DNA sequence, for the purpose of cloned gene integration. The integration rates of a Ty3-neor system (ca. 1 x 10(-3) cell-1 generation-1) and a Ty1-neor system (ca. (2-3) x 10(-3) cell-1 generation-1) were not significantly affected by temperature (18 and 30 degrees C). However, the retrotransposition rate of the Ty3-neor-HIS3 system increased from ca. 2 x 10(-5) to 2 x 10(-4) cell-1 generation-1 when the temperature was decreased from 30 to 18 degrees C. The retrotransposition rate of Ty3-neor was significantly higher than that of Ty3-neor-HIS3 and slightly lower than that of Ty1-neor. This method can be used to estimate integration rates of other Ty3 and Ty1 elements and to evaluate the efficiency of Ty-mediated cloned gene integration.

Cloning, Molecular↗
PubMed1995

Plasmids pIP419 and pIP421 from Bacteroides: 5-nitroimidazole resistance genes and their upstream insertion sequence elements.

The genetic organization of two different 5-nitroimidazole (5-Ni) resistance genes was investigated: nimC and nimD from Bacteroides plasmids pIP419 and pIP421, respectively. The nimC gene (492 bp) and the nimD gene (495 bp) directed the synthesis of polypeptides with deduced molecular masses of 18.37 kDa and 18.48 kDa, respectively. The predicted proteins showed 67-83% identity and 78-91% similarity with the products of two other nimA and nimB genes previously described and could be derived from a common ancestral gene. An insertion sequence element (IS1170) was identified upstream of the nimC gene. IS1170 is 1604 bp in length and is flanked by imperfect inverted repeats (15 bp). IS1170 is similar to the Bacteroides insertion sequence element IS942 with an identity of 70% at the nucleotide level. The single copy of IS1170 present on plasmid pIP419 is integrated 24 bp upstream of the initiation codon of nimC. Similar genetic organization was found on plasmid pIP421. One copy of another insertion sequence (IS1169) was found 4 bp upstream of the first ATG codon of the nimD gene. This element (1325 bp) shows a strong homology at the nucleotide level (70% identity) with IS1186 and IS1168 found to be associated with the Bacteroides carbapenem resistance gene cfiA, and the 5-Nirgenes nimA and nimB, respectively. There is strong evidence that, as in the case of the cfiA gene, the transcription of the four nim genes so far studied is directed by outward-oriented promoters, carried on the right ends of the different insertion sequence elements.

Amino Acid Sequence↗
PubMed2004

Regulated site-specific recombination of the she pathogenicity island of Shigella flexneri.

The she pathogenicity island (PAI) is a chromosomal, laterally acquired, integrative element of Shigella flexneri that carries genes with established or putative roles in virulence. We demonstrate that spontaneous, precise excision of the element from its integration site in the 3' terminus of the pheV tRNA gene is mediated by an integrase gene (int) and a gene designated rox (regulator of excision), both of which are carried on the she PAI. Integrase-mediated excision occurs via recombination between a 22 bp sequence at the 3' terminus of pheV and an imperfect direct repeat at the pheV-distal boundary of the PAI. Excision leads to the formation of a circular episomal form of the PAI, reminiscent of circular excision intermediates of other mobile elements that are substrates for lateral transfer processes such as conjugation, packaging into phage particles and recombinase-mediated integration into the chromosome. The circle junction consists of the pheV-proximal and pheV-distal boundaries of the PAI converging on a sequence identical to 22 bp at the 3' terminus of pheV. The isolated circle was transferred to Escherichia coli where it integrated specifically into phe tRNA genes, as it does in S. flexneri, independently of recA. We also demonstrate that Rox stimulates, but is not essential for, excision of the she PAI in an integrase-dependent manner. However, Rox does not stimulate excision by activating the transcription of the she PAI integrase gene, suggesting that it has an excisionase function similar to that of a related protein from the P4 satellite element of phage P2.

Animals↗
PubMed2006

Multiple genetic elements carry the tetracycline resistance gene tet(W) in the animal pathogen Arcanobacterium pyogenes.

The tet(W) gene is associated with tetracycline resistance in a wide range of bacterial species, including obligately anaerobic rumen bacteria and isolates from the human gut and oral mucosa. However, little is known about how this gene is disseminated and the types of genetic elements it is carried on. We examined tetracycline-resistant isolates of the animal commensal and opportunistic pathogen Arcanobacterium pyogenes, all of which carried tet(W), and identified three genetic elements designated ATE-1, ATE-2, and ATE-3. These elements were found in 25%, 35%, and 60% of tetracycline-resistant isolates, respectively, with some strains carrying both ATE-2 and ATE-3. ATE-1 shows characteristics of a mobilizable transposon, and the tet(W) genes from strains carrying this element can be transferred at low frequencies between A. pyogenes strains. ATE-2 has characteristics of a simple transposon, carrying only the resistance gene and a transposase, while in ATE-3, the tet(W) gene is associated with a streptomycin resistance gene that is 100% identical at the DNA level with the aadE gene from the Campylobacter jejuni plasmid pCG8245. Both ATE-2 and ATE-3 show evidence of being carried on larger genetic elements, but conjugation to other strains was not observed under the conditions tested. ATE-1 was preferentially associated with A. pyogenes strains of bovine origin, while ATE-2 and ATE-3 elements were primarily found in porcine isolates, suggesting that these elements may circulate in different environments. In addition, four alleles of the tet(W) gene, primarily associated with different elements, were detected among A. pyogenes isolates.

Actinomycetaceae↗
PubMed1990

Identification, distribution, and sequence analysis of new insertion elements in Caulobacter crescentus.

We describe two insertion elements isolated from Caulobacter crescentus that are designated IS298 and IS511. These insertion elements were cloned from spontaneous flagellar (fla) gene mutants SC298 and SC511 derived from the wild-type strain CB15 (ATCC 19089), in which they were originally identified as insertions in the flbG operon of the hook gene cluster (N. Ohta, E. Swanson, B. Ely, and A. Newton, J. Bacteriol. 158:897-904, 1984). IS298 and IS511 were each present in C. crescentus CB2 and CB15 in at least four different positions, but neither was present in strain CB13 or in several Caulobacter species examined, including C. vibrioides, C. leidyia, and C. henricii. Nucleotide sequence analysis across the chromosome-insertion element junctions showed that IS298 is located 152 base pairs (bp) upstream from the ATG translation start of the hook protein gene flaK, where it is bounded by a 4-bp direct repeat derived from the site of insertion, and that IS511 is inserted at codon 186 of the flaK coding sequence, where it is also bounded by a 4-bp direct repeat duplicated from the site of insertion. The ilvB102 mutation in strain SC125 was also shown to result from insertion sequence IS511, but no duplication of the genomic sequence was present at the insertion element junctions. IS298 contains an imperfect terminal inverted repeat 16 bp long, and IS511 contains a 32-bp inverted repeat at the termini. IS298 and IS511 are the first insertion elements described in C. crescentus.

Bacteria↗
PubMed1996

A recent fixation of cfiA genes in a monophyletic cluster of Bacteroides fragilis is correlated with the presence of multiple insertion elements.

Small-subunit ribosomal DNA sequences of 16 strains of Bacteroides fragilis were determined and compared with previously published sequences. Three phylogenetic methods (the neighbor-joining, maximum-likelihood, and maximum-parsimony methods) as well as a bootstrap analysis were used to assess the robustness of each topology. All phylogenetic analyses demonstrated that the B. fragilis strains were clearly divided into two robust monophyletic units which corresponded to the cfiA-negative and cfiA-positive groups. Strains of two previously identified DNA homology groups separated similarly into the two monophyletic units. According to the intensity of the hybridization signal with a cfiA probe, the cfiA-positive cluster could be further divided into two groups. This difference might reflect the existence of two, probably closely related cfiA-type genes. In the strongly hybridizing cfiA-positive strains, the gene is capable of conferring high-level resistance to the carbapenems and to most beta-lactamase inhibitors as well, while in the weakly hybridizing cfiA-positive strains, only the latter type of resistance is known to occur. The presence of the cfiA-type genes within a monophyletic cluster of B. fragilis that apparently represents only a minority of the species B. fragilis is suggestive of a recent acquisition. The fact that this cluster is also the predominant pool of all known B. fragilis insertion elements, which have been found to play an important role in the expression of carbapenem resistance, raises the possibility that both genetic determinants, i.e., the resistance gene(s) and insertion elements, may have coevolved.

Bacterial Proteins↗