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H Tobler

Publications and source records attributed to H Tobler.

At least 37 records · Page 2Linked to original sources
PubMed1991

Eliminated chromatin of Ascaris contains a gene that encodes a putative ribosomal protein.

Chromatin diminution in the nematodes Parascaris equorum and Ascaris lumbricoides leads to the formation of somatic cells that contain less DNA than the germ-line cells. We present molecular evidence for the coding potential of germ-line-specific DNA. We report on a cDNA clone that codes for a putative ribosomal protein (ALEP-1, for A. lumbricoides eliminated protein 1). That the corresponding gene is located in the eliminated portion of the genome indicates a difference in germ-line and somatic ribosomes of A. lumbricoides and P. equorum. Elimination of the ALEP-1 gene from all somatic cells in its fully active state may represent an alternative way to gene regulation.

Amino Acid Sequence↗
PubMed1991

Cloning and expression in vitro of a gene encoding tRNAArgACG from the nematode Caenorhabditis elegans.

A gene (rtr-1) coding for the tRNAArgACG has been isolated and characterized from the nematode, Caenorhabditis elegans. The coding portion is not interrupted by an intron and is followed by a track of four thymidines associated with termination by RNA polymerase III. The predicted mature product is 76 nucleotides (nt) long including the CCA tail, and is specific for the most used Arg codon in C. elegans. The gene can be transcribed and processed in a homologous in vitro system. The 82-nt primary transcript begins at the first purine upstream from the mature tRNA 5' end and terminates after the first thymidine of the terminator signal.

Animals↗
PubMed1990

Two highly reiterated nucleotide sequences in the low C-value genome of Panagrellus redivivus.

Two families of highly reiterated satellite nucleotide (nt) sequences have been found in the genome of the sexually separated nematode Panagrellus redivivus. The repeats are arranged in tandem arrays but the different satellites are not intermingled. Monomeric lengths are of 155 bp for one kind and 167 bp for the other; they were named E155 and E167. The A + T content is elevated in both families (i.e., 59.5%, and 65.3%, respectively). No similarity was found between the two satellites nor to other known highly repetitive elements. Furthermore, nt methylation as well as transcriptional activity were negative. An internal subrepeating unit, about 30 bp long, was observed in E167, implying that it could have evolved from a shorter sequence. Reiteration frequencies are approx. 30,000 and 40,000 copies per haploid genome, for E155 and E167, respectively, constituting together about 17% of the total DNA. This figure is astonishingly high, considering a C-value of 70,000 kb in P. redivivus, which is thought to be the lower limit for metazoans. Hence, the genome complexity is approx. 58,000 kb. In contrast to the nematodes Ascaris lumbricoides and Parascaris equorum, however, P. redivivus does not seem to eliminate large blocks of satellite DNA in the presomatic cells during early development.

Animals↗
PubMed1990

Cloning and analysis of three new homeobox genes from the nematode Caenorhabditis elegans.

Three homeobox-containing genes from the nematode Caenorhabditis elegans are described. Two of them (ceh-11 and ceh-12) were isolated from a genomic library by hybridization at low stringency with the Ascaris lumbricoides homeobox AHB-1. The first clone contains a homeobox defining a new class of homeoboxes (ceh-11). This gene maps on the third chromosome of C. elegans, at the same locus as egl-5, a gene already known to be essential for the determination of specific neurons. In the second clone, sequence analysis revealed the existence of the third helix of a putative homeobox (ceh-12) which is interrupted by an intron located upstream of the codon for the amino acid 45 of the homeodomain. Using the ceh-11 homeobox as a probe, a third homeobox (ceh-13) was isolated from a cDNA library. As ceh-13 belongs to the labial class of homeoboxes, we conclude that, at the time when the nematode lineage diverged from the myriapod-insect and the vertebrate lineages, the duplication which led to the Antp and the labial families of homeoboxes had already taken place.

Amino Acid Sequence↗
PubMed1990

Unusual transcription termination of the ribosomal RNA genes in Ascaris lumbricoides.

We studied termination of transcription of the ribosomal RNA genes in Ascaris lumbricoides, the first representative in the phylum of nemathelminthes analysed so far. RNase protection experiments in vivo reveal that the 3' end of the precursor rRNA coincides with the end of mature 26S rRNA. Promoter-containing miniplasmids are able to direct unique 3' end formation in vitro at a site identical to that observed in vivo, whereas deletion of these sequences abolishes 3' end formation throughout the entire spacer. A nuclear run-on experiment in vitro confirms the drop of polymerase I concentration down-stream of this site. The termination site for polymerase I transcription of the rDNA operon in A. lumbricoides is therefore unique, and located at the very end of the 26S rRNA gene.

Animals↗
PubMed1987

Type I-like intervening sequences are found in the rDNA of the nematode Ascaris lumbricoides.

The intervening sequences in the large ribosomal RNA gene of Ascaris lumbricoides var. suum show many similarities to the type I insertions, previously found only in some insect species. They include structural features, but also a presumed transcriptional inactivity in vivo: No transcript of the rDNA intervening sequence in A. lumbricoides could be detected in Northern and dot blot hybridizations. However, the primary structure of the Pol I promoter region is well conserved in interrupted and uninterrupted genes. Moreover, genes with an intervening sequence are correctly initiated in a whole-cell in vitro extract from Ascaris oogonia. Hence, the presence of the intervening sequence alone does not seem to account for a transcriptional inhibition in rRNA genes. As with the type I insertions of insect rDNA, some copies of the A. lumbricoides intervening sequence are also present in locations outside the rDNA cluster. About 50% of the extraribosomal copies are found in a repetitive sequence of the genome, and additional copies are inserted in unique sequences. These striking analogies to type I insertions are discussed, and lead to the conclusion that the two phenomena are undoubtedly related. This is the first report proving the presence of a type I-like insertion element outside of the class Insecta.

Animals↗
PubMed1987

Localization of the in vivo and in vitro transcription initiation site and comparative analysis of the flanking sequences in the two main size classes of Ascaris lumbricoides rDNA.

An accurate in vitro transcription system which utilizes the cloned 8.8 and 8.4 kb size classes of Ascaris rRNA genes (pAlr8 and pAlr13) and two kinds of cellular extracts from Ascaris oogonia has been established. Both rDNA containing plasmids are efficiently transcribed in vitro by RNA polymerase I from a unique site of rDNA which corresponds to the in vivo initiation site. The in vitro transcription product has a triphosphorylated 5'-end and starts on a G localized 414 bp (pAlr8) upstream of the beginning of the mature 18S rRNA. The promoter region has been delimited by testing the in vitro template activity of a series of restriction fragments. The region essential for the accuracy of initiation is contained within nucleotides -72 to +65, but full efficiency of transcription requires the additional presence of the region from nucleotides +66 to +84. The sequences upstream from position -72 do not appear to modulate the efficiency of specific in vitro initiation. Furthermore, the sequences flanking the transcription initiation site from position -1500 to +570 have been determined in the two cloned representatives of the two rDNA main size classes.

Animals↗
PubMed1986

Structure and genomic organization of proretrovirus-like elements partially eliminated from the somatic genome of Ascaris lumbricoides.

A clone containing a middle repetitive element next to satellite DNA has been isolated from a germ line genomic library of the chromatin eliminating nematode Ascaris lumbricoides var. suum. The structure of this element has been elucidated by comparison of several clones containing the element in different environments. It is flanked by 256-bp-long terminal repeats (LTRs) and has an internal region of approximately 7 kb. The nucleotide sequences of both the 5' and the 3' LTRs have been determined. The element has a strong structural similarity with retroviral proviruses and related mobile elements. It was therefore named 'Tas', for transposon-like element of Ascaris. Approximately 50 Tas copies are dispersed over approximately 20 different chromosomal sites. Their genomic distribution varies between individuals, indicating that Tas elements are mobile in the Ascaris genome. Two variant forms, Tas-1 and Tas-2, present in a ratio of approximately 2 to 1 in the germ line genome, have been characterized. They differ not only in their restriction pattern, but also in their elimination behaviour. While only about one-fourth of the Tas-1 elements are expelled from the somatic cell lineage, all Tas-2 copies are specifically eliminated and are thus confined to the germ line cells. We have demonstrated that a cloned representative of Tas-1 elements is expelled concomitantly with its flanking DNA sequences during the chromatin elimination process.

Animals↗
PubMed1986

Characterization of inverted repeated sequences in Ascaris nuclear DNA.

The inverted repeated sequences of the chromatin-eliminating nematode Ascaris lumbricoides var. suum have been examined by electron microscopy and by hydroxyapatite chromatography, both in the germ-line and in the somatic DNA. 38% of the inverted repeats of the germ-line DNA analysed in the electron microscope have a single-stranded loop, in comparison to about 50% of looped structures in the somatic DNA. The loops are on average 2.3 X 10(3) base pairs (bp) long. The rest of the foldback DNA consists of simple hairpins. The average length of looped and unlooped inverted repeats is of the order of 300-400 bp in the germ-line and in the somatic DNA. The content of S1-resistant foldback duplexes isolated by hydroxyapatite chromatography amounts to 1.3% in spermatids, with an average length of 350 bp, and to 1.1% in intestinal or larval cell nuclei, with a length of about 320 bp. We estimate by two different methods that there exist approximately 12500 inverted repeats per haploid germ-line genome and approximately 8000 in the haploid somatic genome. A statistical analysis of the data indicates that the great majority of the foldback sequences are randomly distributed in the Ascaris genome, with a spacing of about (40-80) X 10(3) bp, both in the germ-line and in the somatic DNA.

Animals↗
PubMed1984

Structural organization of the two main rDNA size classes of Ascaris lumbricoides.

The two main rDNA size classes in the genome of Ascaris lumbricoides consist of 8.8 kb and 8.4 kb long repeating units present in a quantitative ratio of roughly 10:1. They both contain the genes coding for 18 , 5.8S and 26S ribosomal RNAs. The length heterogeneity is due to a 450 bp long spacer region localized in the longer repeating unit which begins 870 bp upstream of the 5'-end 18S gene. A few additional microheterogeneities in base sequence occur at the 5'-end of the 26S gene. The 18S, 5.8S and 26S coding regions have been mapped on both the 8.8 kb and 8.4 kb repeating units and the localization of the 5'- and 3'-ends of the 18S and 26S genes has been performed by S1 protection. No intervening sequences are present in either coding region of the two main rDNA size classes.

Animals↗
PubMed1984

Chromosomal arrangement of the two main rDNA size classes of Ascaris lumbricoides.

The two main rDNA size classes of 8.8 kb and 8.4 kb length from Ascaris lumbricoides, which comprise more than 95% of the total nuclear rDNA, are arranged as internally homogeneous clusters and are not intermingled with each other. The rRNA genes are present on a single autosomal locus as could be demonstrated by in situ hybridization of fluorochrome-labelled 18S and 26S rRNA to metaphase chromosomes of spermatocytes I. The quantitative ratio of the two rDNA size classes varies largely between individuals being on the average roughly 10:1 in the investigated wild population of A. lumbricoides. However, there are no differences in the hybridization pattern of the two rDNAs in the DNA eliminating nematode A. lumbricoides between germ line and somatic cells in any of the individuals that have been analyzed.

Animals↗
PubMed1984

Intervening sequences in the ribosomal RNA genes of Ascaris lumbricoides: DNA sequences at junctions and genomic organization.

An rDNA size class in the genome of the nematode Ascaris lumbricoides is described which is interrupted by a 4.5-kb long intervening sequence located in the 26S coding region. This molecular form occurs in approximately 15 copies per haploid genome and amounts to approximately 5% of the total nuclear rDNA. Intervening sequences are present only in the 8.8-kb rDNA, but not in the 8.4-kb rDNA repeating units of A. lumbricoides. Cloning of the interrupted rDNA units revealed, in addition to the main 4.5-kb insertion, shorter intervening sequences of 4-kb and 119-bp length. Both shorter rDNA forms are present in the single copy range of the haploid genome. Sequence analyses of the intervening sequence/rDNA junctions show an identical right-hand junction for all of the three different rDNA forms. The two shorter intervening sequences are a coterminal subset of the right-hand end of the main 4.5-kb insertion, whereas all three insertions have a different left-hand junction with the coding region of rDNA. Each intervening sequence is flanked by a short direct repeat of variable length, being only once present in the uninterrupted rDNA. The intervening sequences of A. lumbricoides show striking similarity to the organization of type I insertion family in dipteran flies, even though they are inserted at different positions in the 26S coding region. Additional rDNA intervening sequences may be present outside of the rDNA cluster, but in not more than 15-20 homologous copies per haploid genome.

Animals↗
PubMed1982

Nucleotide sequence of satellite DNA contained in the eliminated genome of Ascaris lumbricoides.

Several restriction endonuclease fragments isolated from highly repetitive satellite DNA of the chromatin eliminating nematode Ascaris lumbricoides var. suum have been cloned. Each type of restriction fragment corresponds to a different variant of the same related ancestral sequence. These variants differ by small deletions, insertions and single base substitutions. Restriction and DBM blot analyses show that members of the same variant class are tandemly linked and therefore are physically separated from other variant classes. A comparison of all the determined sequences establishes a 121 bp long and AT rich consensus sequence. There is evidence for an internal short range periodicity of 11 bp length, indicating that the Ascaris satellite initially may have evolved from an ancestral undecamer sequence. The satellite DNA sequences are mostly but not entirely eliminated from the presumptive somatic cells during chromatin diminution. We have no evidence for transcriptional activity of satellite DNA at any stage or tissue analyzed.

Animals↗
PubMed1982

Molecular cloning and sequence analysis of highly repetitive DNA sequences contained in the eliminated genome of Ascaris lumbricoides.

High molecular weight DNA from germ line and somatic cells of the DNA eliminating nematode Ascaris lumbricoides has has been isolated and digested with different restriction enzymes. The resulting DNA fragments were separated by agarose gel electrophoresis. Germ line but not somatic DNA shows a prominent band about 120 bp long as well as multiples of that length. These fragments are shown to be monomers and multimers of a highly repetitive satellite DNA, which is eliminated mostly but not completely during the process of chromatin diminution. Restriction digests, hybridization experiments and sequence analysis revealed that this eliminated satellite is composed of a whole set of different but related variant classes, all of them showing the same repeating unit length of about 120 bp. Members of the same variant class are tandemly linked and therefore physically separated from other variant classes. All satellite sequences can be derived from the same common ancestor sequence, differing only by base substitutions, insertions and deletions. There is no evidence for transcription of satellite DNA at any stage and tissues analyzed.

Animals↗
PubMed1978

Quantitative analysis of RNA, glycogen and nucleotides from different developmental stages of Ascaris lumbricoides (var. suum).

Cold acid phenol extractions (Kirby, K.S. (1968) in Methods in Enzymology, Vol. XII B, pp. 87--99, Academic Press, New York) of homogenates from oocytes, zygotes and larval stages of Ascaris lumbricoides yield predominantly glycogen, some glycogen . phosphate complexes and guanine nucleotides, but only small amounts of RNA. Oocytes contain 10.0 pg of extractable 19 S and 26 S rRNA and 1.1 pg 4 S and 5 S RNA per cell. During the maturation of the oocyte to the zygote stage, the amount of 19 S and 26 S rRNA increases by 57%, and that of 4 S and 5 S RNA by 130%. Larval stages contain 62% more 19 S and 26 S rRNA, and 47% more 4 S and 5 S RNA than zygotes. The amount of the extractable glycogen decreases by about 80% and nucleotides by about 70% from oogenesis to the larval stage. The Kirby extract from homogenates of spermatids contains only traces of glycogen, glycogen . phosphate complexes and nucleotides, and 115 times less RNA per spermatid than oocytes. The nucleotide pool sizes of oocytes, zygotes and larval stages were determined; the pools consist of 71--88% guanine nucleotides. Spermatids contain only adenine nucleotides and an unidentified, nucleotide-like compound.

Animals↗