The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated fro... more The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated from Leishmania donovani, and its protein product characterised. It is unique in the Leishmania genome and expressed only in the extracellular promastigote insect form, but not in the intracellular amastigote mammalian form, as shown by northern blots and western blots developed with a specific anti-C terminus immune serum. Indirect immunofluorescence microscopy revealed distinct labelled spots regularly distributed on the plasma membrane, including the part lining the flagellum and the flagellar pocket. By transfection experiments, it was found that wild-type LdARL-3A-overexpressing promastigotes reached higher densities in culture, but released significantly less secreted acid phosphatase in the extracellular medium than the parental strain. When LdARL-3A blocked under the GDP-bound 'inactive' form or with an inactivated potential myristoylation site was overexpressed, the cells d...
Eukaryotic protein-coding genes are generally transcribed by RNA polymerase II (Pol II), which ha... more Eukaryotic protein-coding genes are generally transcribed by RNA polymerase II (Pol II), which has a lower transcription rate than that of Pol I. We report here the duplication of two LD1 genes into the rRNA locus and their resultant transcription by Pol I. The multigenic LD1 locus is present in a 2.2-Mb chromosome in all stocks of Leishmania spp. and is also present in multicopy 200- to 450-kb linear chromosomes or multicopy circular DNAs in over 15% of stocks examined. Genomic rearrangement in Leishmania donovani LSB-51.1 resulted in duplication of a 3.9-kb segment of LD1 containing two genes (orfF and orfG) and of a 1.3-kb segment from approximately 10 kb downstream into the rRNA gene repeat region of the 1.2-Mb chromosome. Short sequences (12 or 13 bp) common to the 2.2-Mb LD1 and 1.2-Mb rRNA loci suggest that this gene conversion occurred by homologous recombination. Transcription of the duplicated genes is alpha-amanitin resistant, indicating transcription by Pol I, in contras...
Ribosomal DNA (rDNA) from three isolates of Trichinella was cloned into phage and sublcloned into... more Ribosomal DNA (rDNA) from three isolates of Trichinella was cloned into phage and sublcloned into the plasmid pBR322. The basic repeat unit of rDNA was variable in size, with the mapped clones ranging from 10-12 kb. There were differences in restriction sites within the genic region among the three isolates which were due to variations in the internal transcribed region (ITS) and the intergenic spacer (IGS). Three RsaI sites were mapped to the IGS repeat unit of the isolate AF1, and one RsaI site was mapped to the IGS repeat unit of Trichinella spiralis pseudospiralis (isolate Tp). The number of repetitive units in the IGS region varied markedly within and between the isolates. It was estimated that the basic repeat unit for the rDNA of isolate P1 was 10.6-28 kb, for AF1 10.7-37 kb, and for Tp it was 11-14.9 kb. There appeared to be a greater frequency of some sizes of the basic repeat unit in each of the populations, based on the relative intensity with which certain bands hybridiz...
Two Trichinella isolates from humans in France were characterized using reproductive capacity ind... more Two Trichinella isolates from humans in France were characterized using reproductive capacity indices and a combination of molecular methods. The isolate TRLL hybridized with the pig type-specific probe pPra and had pig type restriction profiles and rDNA patterns. It was therefore identified as a domestic or pig type isolate. The isolate CTRD-85 had similarities and differences in restriction profiles and rDNA patterns with both AF1 and Trichinella nelsoni and was identified as a sylvatic type. Pattern comparisons also show that T. nelsoni is similar to variants of the North American sylvatic type.
The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated fro... more The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated from Leishmania donovani, and its protein product characterised. It is unique in the Leishmania genome and expressed only in the extracellular promastigote insect form, but not in the intracellular amastigote mammalian form, as shown by northern blots and western blots developed with a specific anti-C terminus immune serum. Indirect immunofluorescence microscopy revealed distinct labelled spots regularly distributed on the plasma membrane, including the part lining the flagellum and the flagellar pocket. By transfection experiments, it was found that wild-type LdARL-3A-overexpressing promastigotes reached higher densities in culture, but released significantly less secreted acid phosphatase in the extracellular medium than the parental strain. When LdARL-3A blocked under the GDP-bound 'inactive' form or with an inactivated potential myristoylation site was overexpressed, the cells d...
Eukaryotic protein-coding genes are generally transcribed by RNA polymerase II (Pol II), which ha... more Eukaryotic protein-coding genes are generally transcribed by RNA polymerase II (Pol II), which has a lower transcription rate than that of Pol I. We report here the duplication of two LD1 genes into the rRNA locus and their resultant transcription by Pol I. The multigenic LD1 locus is present in a 2.2-Mb chromosome in all stocks of Leishmania spp. and is also present in multicopy 200- to 450-kb linear chromosomes or multicopy circular DNAs in over 15% of stocks examined. Genomic rearrangement in Leishmania donovani LSB-51.1 resulted in duplication of a 3.9-kb segment of LD1 containing two genes (orfF and orfG) and of a 1.3-kb segment from approximately 10 kb downstream into the rRNA gene repeat region of the 1.2-Mb chromosome. Short sequences (12 or 13 bp) common to the 2.2-Mb LD1 and 1.2-Mb rRNA loci suggest that this gene conversion occurred by homologous recombination. Transcription of the duplicated genes is alpha-amanitin resistant, indicating transcription by Pol I, in contras...
Ribosomal DNA (rDNA) from three isolates of Trichinella was cloned into phage and sublcloned into... more Ribosomal DNA (rDNA) from three isolates of Trichinella was cloned into phage and sublcloned into the plasmid pBR322. The basic repeat unit of rDNA was variable in size, with the mapped clones ranging from 10-12 kb. There were differences in restriction sites within the genic region among the three isolates which were due to variations in the internal transcribed region (ITS) and the intergenic spacer (IGS). Three RsaI sites were mapped to the IGS repeat unit of the isolate AF1, and one RsaI site was mapped to the IGS repeat unit of Trichinella spiralis pseudospiralis (isolate Tp). The number of repetitive units in the IGS region varied markedly within and between the isolates. It was estimated that the basic repeat unit for the rDNA of isolate P1 was 10.6-28 kb, for AF1 10.7-37 kb, and for Tp it was 11-14.9 kb. There appeared to be a greater frequency of some sizes of the basic repeat unit in each of the populations, based on the relative intensity with which certain bands hybridiz...
Two Trichinella isolates from humans in France were characterized using reproductive capacity ind... more Two Trichinella isolates from humans in France were characterized using reproductive capacity indices and a combination of molecular methods. The isolate TRLL hybridized with the pig type-specific probe pPra and had pig type restriction profiles and rDNA patterns. It was therefore identified as a domestic or pig type isolate. The isolate CTRD-85 had similarities and differences in restriction profiles and rDNA patterns with both AF1 and Trichinella nelsoni and was identified as a sylvatic type. Pattern comparisons also show that T. nelsoni is similar to variants of the North American sylvatic type.
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