Cloning and characterization of β-tubulin genes from Echinococcus multilocularis☆

Brehm, Klaus; Kronthaler, Kerstin; Jura, Heike; Frosch, Matthias

doi:10.1016/s0166-6851(00)00178-x

Cited by 47 publications

(48 citation statements)

References 18 publications

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“…In vitro cultivation of metacestodes was performed according to Jura et al (19), and protoscoleces were isolated as described in Brehm et al (21). Chromosomal DNA and total RNA were isolated from in vivo cultivated parasite material according to previously described protocols (22).…”

Section: Methodsmentioning

confidence: 99%

“…Furthermore, and typical for metazoans, not all echinococcal mRNAs are trans-spliced. Analysis of elp transcripts revealed that trans-spliced and non-trans-spliced mRNAs can even derive from one single gene locus, and recently, we have characterized ␤-tubulin-encoding transcripts of E. multilocularis (21), which are exclusively expressed without spliced leader. Based on a preliminary analysis of 150 randomly chosen, full-length cDNAs (data not shown) we estimate that ϳ25% of all E. multilocularis mRNAs acquire the SL, a figure close to what has been suggested for trematodes (37).…”

Section: Trans-spliced Mrnas Of E Multilocularismentioning

confidence: 99%

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mRNA Trans-splicing in the Human Parasitic CestodeEchinococcus multilocularis

Brehm

Jensen

Frosch

2000

Journal of Biological Chemistry

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An identical 36-nucleotide exon was identified at the 5 termini of different mRNAs from the cestode Echinococcus multilocularis. We provide evidence that this exon constitutes a new spliced leader (SL) that is obviously trans-spliced to echinococcal pre-mRNAs, donated by a non-polyadenylated, trimethylguanosine-capped SL-RNA of 104 nucleotides. Sequence comparisons indicated that cestode and trematode SLs are likely to be derived from a common ancestor gene. No conservation was, however, observed concerning the spectrum of mRNAs that is trans-spliced in cestodes and trematodes, indicating that trans-splicing of a particular flatworm mRNA is not correlated with the function of the encoded protein. We also show that the echinococcal gene elp, encoding a member of the ezrin/radixin/moesin protein family, is expressed via two alternative transcripts, spliced either cis or trans at an identical splice acceptor site. This was accompanied by the formation of different elp primary transcripts, harboring a complete or a truncated upstream intron, which supports the hypothesis that alternative cis/trans-splicing depends on the presence or absence of an upstream splice donor site. A putative SL gene was also identified on chromosomal DNA of Echinococcus granulosus, indicating widespread utilization of trans-splicing in the genus.Trans-splicing is a mechanism of mRNA processing that involves the fusion of exons from independent primary transcripts to form a mature mRNA (for recent reviews, see Refs. 1-5). In the most common form of trans-splicing, called SL 1 trans-splicing, a small "mini-exon" (or SL) is added to the 5Ј ends of pre-mRNA molecules, eventually forming the 5Ј-terminal exon of the mature mRNA. In all cases investigated so far, the SL donor molecule is a small, non-polyadenylated nuclear RNA (the SL-RNA) with structural properties similar to the snRNAs that function in "conventional" cis-splicing (1-5). SL trans-splicing was first described in kinetoplastid protozoans where all mature mRNAs contain an identical leader sequence at the 5Ј end (6). Among metazoans, SL trans-splicing is known for several parasitic and non-parasitic nematodes (1-6) and, in the phylum Platyhelminthes, for parasitic trematodes (7-9) and free-living turbellarians (9). In contrast to kinetoplastids, where trans-splicing is the predominant form of splicing, transcripts of metazoans are generally spliced both cis, usually at introns within the coding region, and trans at the 5Ј end (1-5). A further difference is that in metazoans not all mRNAs are trans-spliced. In Caenorhabditis elegans, for example, around 60% of all transcripts contain one leader, called SL1, and about 10 -15% contain an alternative mini-exon, called SL2 (1-5). Besides SL1 and SL2, three additional SLs were identified in C. elegans that are, however, less frequently found at mRNA 5Ј ends (10).Although significant data has been obtained on the biochemistry of the trans-splicing mechanism itself (1-6), the biological in vivo functions of SLs are less well understood. At ...

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Section: Methodsmentioning

confidence: 99%

Section: Trans-spliced Mrnas Of E Multilocularismentioning

confidence: 99%

mRNA Trans-splicing in the Human Parasitic CestodeEchinococcus multilocularis

Brehm

Jensen

Frosch

2000

Journal of Biological Chemistry

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“…Three isotypes of b-tubulin have been isolated from Caenorhabditis elegans, one of which, mec-7, is more distantly related to the others and forms part of the 15-protofilament tubules, while the other types, Ben 1 and Tub 1, form part of the 11-protofilament microtubules (Savage et al 1989). Three b-tubulin isotypes have also been isolated from Echinococcus multilocularis, one of which (Tub-3) showed a high divergence from the other two; and it has been suggested this too may represent a functionally distinct isotype (Brehm et al 2000). Two small regions of variation occur between the cDNA and the short isotype b-tubulin of G. salaris, in an otherwise relatively conserved region.…”

Section: Discussionmentioning

confidence: 99%

“…If one BZ-susceptible isotype is expressed in a parasite, even if other resistant isotypes are also present, it is likely that there will be an overall sensitivity to BZ drugs (Brehm et al 2000). Nevertheless, BZ-resistant isotypes can become dominant in a population through selection (Kwa et al 1993a).…”

Section: Discussionmentioning

confidence: 99%

“…The b-tubulin gene of helminths has been found to contain a number of introns, with the exception of that isolated from Fasciola hepatica, which vary greatly in sequence between species (Brehm et al 2000;Robinson et al 2001), offering the possibility of their use as diagnostic markers. The classic method of Gyrodactylus identification relies on morphological characteristics of the attachment organ of the parasite (Malmberg 1970); and it can be difficult to distinguish between closely related species using this method.…”

Section: Introductionmentioning

confidence: 99%

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Characterisation of a �-tubulin gene from the monogenean parasite, Gyrodactylus salaris Malmberg, 1957

Collins

Miller

Cunningham

2004

Parasitology Research

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The cDNA and two partial genomic sequences of beta-tubulin genes have been isolated from the monogenean parasite Gyrodactylus salaris. The cDNA sequence is not represented by either of the genomic sequences, implying that at least three isotypes of the gene exist in G. salaris. The sequences show regions of high homology with other helminth beta-tubulin genes. This represents the first isolation of a beta-tubulin gene from a monogenean and contributes to the overall characterisation of these genes within the helminths. This is an important area, as anthelmintic resistance is increasing against benzimidazole drugs that target the beta-tubulin gene. Benzimidazole drugs have been tested successfully against Gyrodactylus parasites, but their use is not widespread. Should it increase, analysis of the beta-tubulin gene may provide a tool for monitoring resistance development and improving management practises. Use of the beta-tubulin gene in the identification of Gyrodactylus species may prove complex due to the presence of different isotypes.

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Protein Kinases as Drug Targets in the Treatment of Alveolar Echinococcosis

Brehm

2013

Protein Phosphorylation in Parasites

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Cloning and characterization of β-tubulin genes from Echinococcus multilocularis☆

Cited by 47 publications

References 18 publications

mRNA Trans-splicing in the Human Parasitic CestodeEchinococcus multilocularis

mRNA Trans-splicing in the Human Parasitic CestodeEchinococcus multilocularis

Characterisation of a �-tubulin gene from the monogenean parasite, Gyrodactylus salaris Malmberg, 1957

Protein Kinases as Drug Targets in the Treatment of Alveolar Echinococcosis

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