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T Langin

Publications and source records attributed to T Langin.

At least 37 records · Page 2Linked to original sources

Fot 1 insertions in the Fusarium oxysporum f. sp. albedinis genome provide diagnostic PCR targets for detection of the date palm pathogen.

Populations of Fusarium oxysporum f. sp. albedinis, the causal agent of Bayoud disease of date palm, are derivatives of a single clonal lineage and exhibit very similar Fot 1 hybridization patterns. In order to develop a sensitive diagnostic tool for F. oxysporum f. sp. albedinis detection, we isolated several DNA clones containing a copy of the transposable element Fot 1 from a genomic library of the date palm pathogen. Regions flanking the insertion sites were sequenced, and these sequences were used to design PCR primers that amplify the DNA regions at several Fot 1 insertion sites. When tested on a large sample of Fusarium isolates, including 286 F. oxysporum f. sp. albedinis isolates, 17 other special forms, nonpathogenic F. oxysporum isolates from palm grove soils, and 8 other Fusarium species, the primer pair TL3-FOA28 allowed amplification of a 400-bp fragment found only in F. oxysporum f. sp. albedinis. Sequence analysis showed that one of the Fot 1 copies was truncated, lacking 182 bp at its 3' terminus. The primer pair BI03-FOA1 amplified a 204-bp fragment which overlapped the Fot 1 truncated copy and its 3' site of insertion in the F. oxysporum f. sp. albedinis genome and identified 95% of the isolates. The primer pairs BIO3-FOA1 and TL3-FOA28 used in PCR assays thus provide a useful diagnostic tool for F. oxysporum f. sp. albedinis isolates.

Cloning, Molecular↗

Upper airway changes in snorers and mild sleep apnea sufferers after uvulopalatopharyngoplasty (UPPP).

OBJECTIVES: We used upper airway (UA) imaging in 20 patients to determine (1) whether an effective enlargement of the UA is obtained after uvulopalatopharyngoplasty (UPPP), and (2) whether UA modifications explain the results of such surgery. METHODS: Cephalometric measurements were made to assess the posterior airway space, the length and width of the soft palate, and the distance between the hyoid bone and the mandibular plane. Pharyngeal CT measured the airway cross-sectional area (CSA) at each 10-mm slice from 10 mm above (-10) to 40 mm below (+40) the hard palate. Polysomnography was performed before and after surgery (10+/-10 [SD] months). Good responders were defined by an apnea-hypopnea index (AHI) of <10 postsurgery or, in patients in whom AHI was initially <10, a reduction of AHI >50% of the initial AHI. RESULTS: Twenty patients (age=45+/-11 years) were studied. For the whole group, the mean body mass index (26+/-4 kg/m2) and AHI (14+/-13 vs 18+/-16/h) were unchanged after UPPP. The results of the surgery were mediocre with 7 good responders (35%) and 13 nonresponders (65%) defined by polysomnographic criteria. The only changes on UA imaging for the group as a whole after UPPP were decrease in length (40+/-6 vs 29+/-5 mm, p< or =0.0006) and increase in width of the soft palate (11.5+/-2.7 vs 13.6+/-3.5 mm, p< or =0.006). The increase or decrease in minimal CSA at the oropharyngeal (OP) level after UPPP was significantly correlated with the change in AHI (r=-0.54, p<0.02). Moreover, the changes in CSA obtained at the OP level were significant only in the patients who responded favorably to UPPP (7 vs 13 nonresponders). CONCLUSIONS: Postoperative OP enlargement is associated with a good outcome of UPPP. Persistent narrowing in nonresponders could be due to the increase in soft palate width after surgery.

Adult↗

[Acute eosinophilic pneumonia due to hypersensitivity to carbamazepine].

The authors report a case of hypersensitivity to carbamazepine presenting as an acute eosinophilic pneumonia and a probable hypersensitivity myopericarditis in a 54-year-old patient treated with carbamazepine for three weeks. Treatment consisted of stopping the carbamazepine and in prolonged corticosteroid therapy. The features of the hypersensitivity syndrome to carbamazepine are mentioned as well as the different pulmonary disorders attributable to carbamazepine.

Adrenal Cortex Hormones↗

Isolation of the transposable element hupfer from the entomopathogenic fungus Beauveria bassiana by insertion mutagenesis of the nitrate reductase structural gene.

A transposable element has been isolated from the entomopathogenic fungus Beauveria bassiana by trapping it in the nitrate reductase structural gene, which has been cloned from this species. The element had inserted in the first exon of the nia gene and appeared to have duplicated the sequence TA at the site of insertion. It was 3336 bp long with 30-bp imperfect, inverted, terminal repeats. The element, called hupfer, contained an open reading frame encoding a 321-amino acid protein similar to the IS630- or mariner-Tcl-like transposases, and a residual sequence of about 2 kb which was not significantly similar to any published sequence. There are fewer than five copies of this transposable element present per genome in the fungus.

Amino Acid Sequence↗

Do the integrases of LTR-retrotransposons and class II element transposases have a common ancestor?

The integrases of retrotransposons (class I) and retroviruses and the transposases of bacterial type elements (class II) were compared. The DDE signature that is crucial for the integration of these elements is present in most of them, except for the non-LTR retrotransposons and members of the hAT and P super-families. Alignment of this region was used to infer the relationships between class II elements, retrotransposons, and retroviruses. The mariner-Tc1 and the Pogo-Fot1 super-families were found to be closely related and probably monophyletic, as were LTR retrotransposons and retroviruses. The IS elements of bacteria were clustered in several families, some of them being closely related to the transposase of the mariner-Tc1 super-family or to the LTR retrotransposon and retrovirus integrases. These results plus that of Xiong and Eickbush (1990) were used to develop an evolutionary history suggesting a common ancestral origin(s) for the integrases and transposases containing the DDE signature. The position of the telomeric elements (Het-A and TART) was assessed by comparing their gag and reverse transcriptase domains (when present) to those of group II introns and non-LTR retrotransposons. This preliminary analysis suggests that telomeric elements may be derived from non-LTR retrotransposons.

Amino Acid Sequence↗

Relationships between transposable elements based upon the integrase-transposase domains: is there a common ancestor?

The integrase domain of RNA-mediated elements (class I) and the transposase domain of DNA-mediated transposable elements (class II) were compared. A number of elements contain the DDE signature, which plays an important role in their integration. The possible relationships between mariner-Tc1 and IS elements, retrotransposons, and retroviruses were analyzed from an alignment of this region. The mariner-Tc1 superfamily, and LTR retrotransposons and retroviruses were found to be monophyletic groups. However, the IS elements of bacteria were found in several groups. These results were used to propose an evolutionary history that suggests a common ancestor for some integrases and transposases.

Amino Acid Sequence↗

[Endobronchial stenoses in Wegener's granulomatosis].

The authors describe a case of a patient suffering from Wegener's disease with endobronchial manifestations which progressively developed tracheal stenosis which was not accessible to the usual immunodepressive therapy. Repeated endobronchial dilatation using a balloon introduced via supple fibroscope was carried out with success. The balloon dilatation seems a useful alternative approach to other therapies such as surgery. It is a non-aggressive technique which is easily performed and has no complications.

Biopsy↗

The transposable element impala, a fungal member of the Tc1-mariner superfamily.

A new transposable element has been isolated from an unstable niaD mutant of the fungus Fusarium oxysporum. This element, called impala, is 1280 nucleotides long and has inverted repeats of 27 bp. Impala inserts into a TA site and leaves behind a "foot-print" when it excises. The inserted element, impala-160, is cis-active, but is probably trans-defective owing to several stop codons and frameshifts. Similarities exist between the inverted repeats of impala and those of transposons belonging to the widely dispersed mariner and Tc1 families. Moreover, translation of the open reading frame revealed three regions showing high similarities with Tc1 from Caenorhabditis elegans and with the mariner element of Drosophila mauritiana. The overall comparison shows that impala occupies an intermediate position between the mariner and Tc1-like elements, suggesting that all these elements belong to the same superfamily. The degree of relatedness observed between these elements, described in different kingdoms, raises the question of their origin and evolution.

Amino Acid Sequence↗

Horizontal transmission versus ancient origin: mariner in the witness box.

The transposable element mariner has been found in many species of Drosophilidae, several groups of Arthropods, and more recently in Platyhelminthes as well as in a phytopathogenic fungus. In the family Drosophilidae, the distribution of mariner among species shows many gaps, and its geographical distribution among endemic species is restricted to Asia and Africa. Among mariner elements in species within and outside the Drosophilidae, the similarities in nucleotide sequence and the amino acid sequence of the putative transposase reveal many phylogenetic inconsistencies compared with the conventional phylogeny of the host species. This paper discusses the contrasting hypotheses of horizontal transfer versus ancestral origin proposed to explain these results.

Africa↗

The strange phylogenies of transposable elements: are horizontal transfers the only explantation?

Analyses of the evolution of transposable elements reveal some inconsistencies when the phylogenies of such elements are compared to conventional phylogenies of the host species. Such discrepancies are generally interpreted as resulting from occasional horizontal transfers of transposable elements across species boundaries. This phenomenon has been clearly demonstrated for only a few elements and both its frequency and the mechanism by which it occurs remain unknown. Moreover, in many cases, the hypothesis of horizontal transfer must be compared with alternative evolutionary scenarios.

Animals↗

The nia gene of Fusarium oxysporum: isolation, sequence and development of a homologous transformation system.

The Fusarium oxysporum gene nia, encoding nitrate reductase (NR), was isolated from a cosmid library by direct complementation of an F. oxysporum nia- mutant. The gene specifies a protein of 905 amino acids and contains a 57-bp intron. Comparison of the deduced aa sequence with NR of other fungi revealed a high degree of similarity and conservation in the intron position. The cloned nia made it possible to develop the first homologous transformation system for this fungus. Transformation frequencies of up to 600 transformants per microgram of DNA were achieved. Gene replacement, single-copy homologous integrations and integrations at non-homologous sites were observed. Direct comparison between plasmids and cosmids carrying the same gene showed a higher frequency of targeted transformation using cosmid vectors. Gene replacement events were observed in about 50% of the transformants analysed with each type of vector used. This high frequency of substitution offers new applications for the transformation system in F. oxysporum.

Amino Acid Sequence↗

The GUS gene fusion system (Escherichia coli beta-D-glucuronidase gene), a useful tool in studies of root colonization by Fusarium oxysporum.

The plant-pathogenic fungus Fusarium oxysporum was successfully transformed with the beta-D-glucuronidase gene from Escherichia coli (gusA) (GUS system) in combination with the gene for nitrate reductase (niaD) as the selectable marker. The frequency of cotransformation, as determined by GUS expression on plates containing medium supplemented with 5-bromo-4-chloro-3-indolyl glucuronide (GUS+), was very high (up to 75%). Southern hybridization analyses of GUS+ transformants revealed that single or multiple copies of the gusA gene were integrated into the genomes. High levels of GUS activity are expressed in some transformants, but activity in F. oxysporum does not appear to be correlated with the copy number of the gusA gene. Since the highest activity was found in a transformant with a single copy, it can be assumed that sequence elements of F. oxysporum integrated upstream of the gene can act as a promoter or enhancer. Expression of the gusA gene was also detected during growth of the fungus in plants, indicating that the GUS system can be used as a sensitive and easy reporter gene assay in F. oxysporum.

Cloning, Molecular↗

Fot1, a new family of fungal transposable elements.

We report here the discovery of a family of transposable elements, which we refer to as Fot1 elements, in the fungal plant pathogen Fusarium oxysporum. The first element was identified as an insertion in the gene encoding nitrate reductase. It is 1928 bp long, has 44 bp inverted terminal repeats, contains a large open reading frame and is flanked by a 2 bp (TA) target site duplication. This element shares significant structural similarities with a class of transposons that includes Tc1 from Caenorhabditis elegans and therefore represents a new class of transposable elements in fungi.

Amino Acid Sequence↗

Heterologous expression of the Aspergillus nidulans regulatory gene nirA in Fusarium oxysporum.

We have isolated strains of Fusarium oxysporum carrying mutations conferring a phenotype characteristic of a loss of function in the regulatory gene of nitrate assimilation (nirA in Aspergillus nidulans, nit-4 in Neurospora crassa). One of these nir- mutants was successfully transformed with a plasmid containing the nirA gene of A. nidulans. The nitrate reductase of the transformants is still inducible, although the maximum activity is lower than in the wild type. Single and multiple integration events were found, as well as a strict correlation between the presence of the nirA gene and the Nir+ phenotype of the F. oxysporum transformants. We also investigated how the A. nidulans structural gene (niaD) is regulated in F. oxysporum. Enzyme assays and Northern experiments show that the niaD gene is subject to nitrate induction and that it responds to nitrogen metabolite repression in a F. oxysporum genetic background. This indicates that both the mechanisms of specific induction, mediated by a gene product isofunctional to nirA, and nitrogen metabolite repression, presumably mediated by a gene product isofunctional to the homologous gene of A. nidulans, are operative in F. oxysporum.

Aspergillus nidulans↗

Reciprocal exchanges instigated by large heterologies in the b2 gene of ascobolus are not associated with long adjacent hybrid DNA stretches.

In the gene b2 of Ascobolus immersus, large heterologies increase the frequencies of reciprocal exchanges on their upstream border (corresponding to the high non-Mendelian segregation side). Tests were made to determine whether these reciprocal exchanges, instigated by large heterologies, resulted from the blockage of a Holliday junction bordering a hybrid DNA tract extending from the end of the gene to the heterology. Three types of experiments were performed to answer this question. In all cases, results did not correlate the presence of reciprocal exchanges instigated by large heterologies with the presence of adjacent hybrid DNA tracts. These reciprocal exchanges were rarely associated with postmeiotic segregation at upstream markers, they were not associated with gene conversion of a marker within the interval and their frequency was not decreased by decreasing the frequency of hybrid DNA formation in the gene. These results led to the proposal of the existence of a precursor to reciprocal exchange different from a single branch-migrating Holliday junction. This precursor migrates rightward and its migration is dependent on the DNA sequence homology. The existence of this precursor does not exclude that reciprocal exchanges resulting from the maturation of single Holliday junctions bordering adjacent hybrid DNA tracts could also occur.

Ascomycota↗

Hybrid DNA extension and reciprocal exchanges: alternative issues of an early intermediate during meiotic recombination?

Large heterologies in gene b2 strongly increase the frequencies of reciprocal exchanges on their left border, towards the high conversion end. In a previous study, we observed that heterozygous point mutations located in the high conversion end (region F) stimulate the reciprocal exchanges instigated by the large heterology 138. We have defined some properties of this stimulation. The effect does not depend on the nature of the large heterology used. It is effective only with point mutations located on the left side of the large heterology. It does not depend on the number of heterozygosities accumulated in region F. It is not specific on the location of point mutations in region F: it decreases from region F (left end) to region E (middle part of b2). It is correlated with the mismatch correction efficiencies of the point mutations used. It is not observed in the absence of a large heterology. Point mutation heterozygosities which stimulate reciprocal exchanges also decrease the frequency of HDNA formation in gene b2. We propose a model in which reciprocal exchanges on the one hand and hybrid DNA formation on the other hand correspond to alternative processings of a common recombination intermediate.

Ascomycota↗

The MET2 gene of Saccharomyces cerevisiae: molecular cloning and nucleotide sequence.

A 5.1-kb DNA fragment from Saccharomyces cerevisiae, which complements a yeast met2 mutant strain, has been cloned. This fragment contains the wild-type MET2 gene which codes for the homoserine O-transacetylase, one of the methionine biosynthetic enzymes. The presence of the MET2 gene has been shown by integrative transformation experiments and genetic analyses of the resulting transformants. The complete nucleotide sequence of a 2826-bp DNA fragment carrying the MET2 gene has been determined. The sequence contains one major open reading frame of 438 codons, giving a calculated Mr of 48,370 for the encoded protein. We have identified the transcriptional product of the MET2 gene and estimated its size at 1650 nucleotides.

Acetyltransferases↗