Sequence comparison of a segment of the gene for 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase in zygomycetes

Burmester, Anke; Czempinski, Katrin

doi:10.1111/j.1432-1033.1994.tb18637.x

Cited by 26 publications

(20 citation statements)

References 26 publications

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“…The hybridization conditions for DNA were as described by Burmester & Czempinski (1994). A digoxigenin-labelled probe of a partial sequence of sexP and sexM was used, amplified with primer pairs M and N (Supplementary Table S1).…”

Section: Methodsmentioning

confidence: 99%

The mating-related loci sexM and sexP of the zygomycetous fungus Mucor mucedo and their transcriptional regulation by trisporoid pheromones

et al. 2012

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The putative mating type locus of mucoralean fungi consists of a single high mobility group (HMG)-domain transcription factor gene, sexM or sexP, flanked by genes for an RNA helicase and a triosephosphate transporter. We used degenerate primers derived from the amino acid sequence of the RNA helicase to sequence a fragment of this gene from Mucor mucedo. This fragment was extended by inverse PCR to obtain the complete sequences of the sex loci from both mating types of M. mucedo. The sex loci in M. mucedo reflect the general picture obtained previously for Phycomyces blakesleeanus, presenting a single HMG-domain transcription factor gene, sexM and sexP in the minus and plus mating types, respectively. These are located next to a gene for RNA helicase. Transcriptional analysis by quantitative real-time PCR showed that only transcription of sexM is considerably stimulated by adding trisporoid pheromones, thus mimicking sexual stimulation, whereas sexP is only slightly affected. These differences in regulation between sexM and sexP are supported by the observation that the promoter sequences controlling these genes show no similarities. The protein structures themselves are considerably different. The SexM, but not the SexP protein harbours a nuclear localization sequence. The SexM protein is indeed transported to nuclei. This was shown by means of a GFP fusion construct that was used to study the localization of SexM in the yeast Saccharomyces cerevisiae. The fusion protein is highly enriched in nuclei.

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Section: Methodsmentioning

confidence: 99%

The mating-related loci sexM and sexP of the zygomycetous fungus Mucor mucedo and their transcriptional regulation by trisporoid pheromones

et al. 2012

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“…A cloned HindIII-SalI fragment was used as the probe for a DNA-region containing a putative suppressor of temperature-sensitive DNA polymerase a mutations (PSP1; Formosa & Nittis, 1998). Hybridization and detection conditions were as described by Burmester & Czempinski (1994), and DNA sequencing was performed by Eurofins MWG-Operon.…”

Section: Methodsmentioning

confidence: 99%

Complementation of a stable Met2-1 mutant of the zygomycete Absidia glauca by the corresponding wild-type allele of the mycoparasite Parasitella parasitica, transferred during infection

et al. 2013

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Compared with prokaryotes, where horizontal gene transfer events are frequently found and can be studied in the laboratory at the mechanistic level, few systems are known that allow direct experimental access to parasexual phenomena in eukaryotes. In zygomycetes, a basal lineage of fungi, several mycoparasitic fungi are known that inevitably form a cytoplasmic continuum with their hosts during infection. We provide evidence that, corresponding to the expectation suggested by the morphology of the infection process, gene transfer occurs from the parasite to the host. For analysing this parasexual system at the DNA level, we characterized interspecific recombinants obtained by infecting a stable methionine-auxotrophic Absidia glauca mutant with heavy rearrangements at the Met2-1 locus, which encodes homoserine acetyltransferase. Recipients were shown to be complemented by part of the corresponding gene from Parasitella parasitica. This foreign DNA is neither integrated at the putative Met2-2 locus in the recipient strain nor integrated at Met2-1, a locus encoding a hypothetical protein with amino acid similarity but with unknown function. Based on hybridization studies and on the phenotype of recipients that bear some mitotic instability of the acquired prototrophy, we propose that P. parasitica DNA is established in A. glauca recipients as extrachromosomally located replicons.

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“…Electroporation parameters were: 400 V, 25 µF for 0.4 cm cuvettes using the Biorad gene pulser apparatus. For analysis of transformants by Southern blot analysis, stringent hybridization and washing conditions were used (Burmester and Czempinski 1994). Hybridization probes were prepared by labeling a plasmid containing the gfp gene pTrcHisXpress (InVitrogen) by the method of Feinberg and Vogelstein (1984).…”

Section: Transformation and Southern Blottingmentioning

confidence: 99%

Green fluorescent protein as a reporter for gene expression in the mucoralean fungus Absidia glauca

Schilde

Wöstemeyer

Burmester

2001

Archives of Microbiology

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Mucoralean fungi (Zygomycota) are used for many industrial processes and also as important model organisms for investigating basic biological problems. Their genetic analysis is severely hampered by low transformation frequencies, by their strong tendency towards autonomous replication of plasmids instead of stable integration, and by the lack of reliable genetic reporter systems. We constructed plasmids for transforming the model zygomycete Absidia glauca that carry the versatile reporter gene coding for green fluorescent protein (GFP). gfp expression is controlled either by the homologous actin promoter or the promoter for the elongation factor of translation, EF1alpha. These plasmids also confer neomycin resistance and carry one of two genetic elements (rag1, seg1) that improve mitotic stability of the plasmid. The gfp constructs were replicated extrachromosomally and could be recovered from retransformed Escherichia coli cells. gfp expression was monitored by epifluorescence microscopy. The gfp reporter gene plasmids presented here for the model zygomycete A. glauca constitute the first reliable system that allows the monitoring of gene expression in this important group of fungi.

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Sequence comparison of a segment of the gene for 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase in zygomycetes

Cited by 26 publications

References 26 publications

The mating-related loci sexM and sexP of the zygomycetous fungus Mucor mucedo and their transcriptional regulation by trisporoid pheromones

The mating-related loci sexM and sexP of the zygomycetous fungus Mucor mucedo and their transcriptional regulation by trisporoid pheromones

Complementation of a stable Met2-1 mutant of the zygomycete Absidia glauca by the corresponding wild-type allele of the mycoparasite Parasitella parasitica, transferred during infection

Green fluorescent protein as a reporter for gene expression in the mucoralean fungus Absidia glauca

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