A Unique Repeated DNA Sequence in the Cyclosporin-Producing Strain of Tolypocladium inflatum (ATCC 34921)

Kempken, Frank; Schreiner, Claudia; Schörgendorfer, Kurt; Kück, Ulrich

doi:10.1006/emyc.1995.1037

Cited by 17 publications

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“…This method allowed the differentiation of clones carrying single-or multicopy sequences (24). 32 P-labeled yeast rDNA (pMY60 [59]) was used to identify clones containing T. inflatum rDNA sequences (34). About 1% of 4,800 plaques tested contained repeated DNA sequences exhibiting no rDNA sequence homology.…”

Section: Resultsmentioning

confidence: 99%

“…All seven chromosomes of T. inflatum (54) hybridized with the probe, indicating that the RE is indeed present on all chromosomes. A more detailed view of the RE's genomic distribution was obtained via an analysis of chromosomal DNA restriction fragments (34,35). T. inflatum chromosomes were digested with rare-cutting enzymes, and the fragments were then separated by pulsed-field gel electrophoresis, Southern blotted, and probed with the RE (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Chromosomal DNA was isolated and run on pulsed-field gels as described elsewhere (54). A restriction analysis of high-molecular-weight chromosomal DNA was undertaken, using rare-cutting endonucleases in a previously described method (34,35).…”

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confidence: 99%

“…In an attempt to circumvent this problem, electrophoretic karyotypes of both T. inflatum and its related strains were established (54). In addition, a strain-specific unique repeated DNA element (CPA) was also characterized (34). To date, isolation of genes involved in regulating cyclosporine synthase gene expression has been unsuccessful, largely because of limitations in standard molecular genetic approaches.…”

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restless, an Active Ac-like Transposon from the Fungus Tolypocladium inflatum: Structure, Expression, and Alternative RNA Splicing

Kempken

Kück

1996

Molecular and Cellular Biology

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Elements of the hAT transposon family, such as the maize activator (Ac), have been discovered in a large number of eukaryotic species. This type of class II transposon, present in both plants and animals, has not been previously detected in any fungal organism. However, using a differential screening method to detect repetitive DNA, we have identified a hAT transposon in the hyphomycete Tolypocladium inflatum. The transposon, which we named restless, is 4,097 bp long, carries 20-bp inverted repeats and an 8-bp target site duplication, and encodes a long open reading frame which is interrupted by a single intronic sequence. The derived mRNA exhibits alternative splicing, resulting in the formation of two transcripts that may be translated into polypeptides of 157 or 803 amino acids. The predicted amino acid sequence of the larger polypeptide demonstrates significant homology with transposases from the hAT transposon family. A chromosomal analysis using pulsed-field gel electrophoresis showed that all seven chromosomal bands carry copies of the 4.1-kb transposon. This was confirmed in hybridization experiments with rare-cutting restriction endonucleases which indicate that about 15 copies are present in T. inflatum. The genomic distribution of restless and its transcriptional expression, alternative mRNA splicing, and genomic mobility all imply a potential role for this element in developing a transposon tagging system for use in filamentous fungi.Eukaryotic transposons can be divided into two major groups (18, 42). Class I transposable elements are able to transpose via an RNA intermediate, a process which employs a reverse transcriptase. Well-known members of this type include retrotransposons such as the copia element from Drosophila melanogaster (17) and the Ty elements from Saccharomyces cerevisiae (61).Class II elements, which transpose via DNA intermediates, demonstrate direct excision prior to their integration into target sequences. The best-known examples are the Ac-Ds elements of maize, discovered about half a century ago by Barbara McClintock (40,41).Both classes of transposons have previously been identified in filamentous fungi. In a few cases, such as the Neurospora crassa Tad element (37) and the Fusarium oxysporum Fot1 and impala elements (10, 39), this was achieved by the use of a transposon trap. Others were discovered by characterizing repeated DNA sequences, leading to the isolation of the MGR (24) and grasshopper (13) elements from Magnaporthe grisea, the Foret element from F. oxysporum (31), and the CfT-1 element from Cladosporium fulvum (43). The majority of these fungal transposons are class I transposable elements, while only a few belong to class II (Fot1 and impala). Those elements, whose prototypes were found in Caenorhabditis elegans or in D. melanogaster (7,28,46), were characterized as Tc1-mariner-like transposons (9).Here, we describe the isolation and characterization of a class II transposon from Tolypocladium inflatum ATCC 34921 (synonym Beauveria nivea [20,60]). This filamentou...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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restless, an Active Ac-like Transposon from the Fungus Tolypocladium inflatum: Structure, Expression, and Alternative RNA Splicing

Kempken

Kück

1996

Molecular and Cellular Biology

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“…Interestingly, politons share some characteristics with mitochondrial linear plasmids [10,11] There are some fungal repetitive elements that have not been characterized in detail. Several years ago, a repetitive sequence was discovered in the genome of the filamentous fungus, Tolypocladium inflatum; this element was called CPA [14]. When first detected it was found to be strain-specific, but no evidence supporting its inclusion with either class I or class II transposons was apparent.…”

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The Tolypocladium inflatum CPA element encodes a RecQ helicase‐like gene

Kempken

2008

J. Basic Microbiol.

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Previously, a repetitive CPA element was discovered in the genome of the filamentous fungus Tolypocladium inflatum; however, no further characterization was technically possible at that time. In this study, PCR amplification was used to detect a 4 kb conserved portion of the CPA element that appeared to be present in most, if not all, genomic CPA elements. The amplicons included a large open reading frame that was most similar to a RecQ helicase-like gene from Metarhizium anisopliae. The repetitive nature of the CPA element suggests that it is related to the eukaryotic Helitron class of transposable elements.

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Transposons in filamentous fungi-facts and perspectives

1998

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Transposons are ubiquitous genetic elements discovered so far in all investigated prokaryotes and eukaryotes. In remarkable contrast to all other genes, transposable elements are able to move to new locations within their host genomes. Transposition of transposons into coding sequences and their initiation of chromosome rearrangements have tremendous impact on gene expression and genome evolution. While transposons have long been known in bacteria, plants, and animals, only in recent years has there been a significant increase in the number of transposable elements discovered in filamentous fungi. Like those of other eukaryotes, each fungal transposable element is either of class I or of class II. While class I elements transpose by a RNA intermediate and employ reverse transcriptases, class II elements transpose directly at the DNA level. We present structural and functional features for such transposons that have been identified so far in filamentous fungi. Emphasis is given to specific advantages or unique features when fungal systems are used to study transposable elements, e.g., the evolutionary impact of transposons in coenocytic organisms and possible experimental approaches toward horizontal gene transfer. Finally, we focus on the potential of transposons for tagging and identifying fungal genes.

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A Unique Repeated DNA Sequence in the Cyclosporin-Producing Strain of Tolypocladium inflatum (ATCC 34921)

Cited by 17 publications

References 0 publications

restless, an Active Ac-like Transposon from the Fungus Tolypocladium inflatum: Structure, Expression, and Alternative RNA Splicing

restless, an Active Ac-like Transposon from the Fungus Tolypocladium inflatum: Structure, Expression, and Alternative RNA Splicing

The Tolypocladium inflatum CPA element encodes a RecQ helicase‐like gene

Transposons in filamentous fungi-facts and perspectives

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