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R Erdmann

Publications and source records attributed to R Erdmann.

47 records · Page 3Linked to original sources
PubMed1992

Peroxisomal multifunctional beta-oxidation protein of Saccharomyces cerevisiae. Molecular analysis of the fox2 gene and gene product.

The gene encoding the multifunctional protein (MFP) of peroxisomal beta-oxidation in Saccharomyces cerevisiae was isolated from a genomic library via functional complementation of a fox2 mutant strain. The open reading frame consists of 2700 base pairs encoding a protein of 900 amino acids. The predicted molecular weight (98,759) is in close agreement with that of the isolated polypeptide (96,000). Analysis of the deduced amino acid sequence revealed similarity to the MFPs of two other fungi but not to that of rat peroxisomes or the multifunctional subunit of the Escherichia coli beta-oxidation complex. The FOX2 gene was overexpressed from a multicopy vector (YEp352) in S. cerevisiae and the gene product purified to apparent homogeneity. A truncated version of MFP lacking 271 carboxyl-terminal amino acids was also overexpressed and purified. Experiments to study the enzymatic properties of the wild-type MFP demonstrated an absence of activities originally assigned to an MFP of S. cerevisiae (crotonase, L-3-hydroxyacyl-CoA dehydrogenase, and 3-hydroxyacyl-CoA epimerase), whereas two other activities were found: 2-enoyl-CoA hydratase 2 (converting trans-2-enoyl-CoA to D-3-hydroxyacyl-CoA) and D-3-hydroxyacyl CoA dehydrogenase (converting D-3-hydroxyacyl-CoA to 3-ketoacyl-CoA). The truncated form contained only the D-3-hydroxyacyl-CoA dehydrogenase activity. These results clearly demonstrate that the beta-oxidation of fatty acids in S. cerevisiae follows a previously unknown stereochemical course, namely it occurs via a D-3-hydroxyacyl-CoA intermediate.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1991

Regulation of transcription of the gene coding for peroxisomal 3-oxoacyl-CoA thiolase of Saccharomyces cerevisiae.

Transferring Saccharomyces cerevisiae cells from glucose- to oleate-containing growth media results in a significant increase in the number and volume of peroxisomes. To investigate this proliferation process we studied the transcriptional regulation of the gene coding for peroxisomal 3-oxoacyl-CoA thiolase (EC 2.3.1.16) in response to the switch in carbon source. Expression was proved to be repressed during growth on glucose, derepressed during growth on glycerol, and induced during growth on oleate as the sole carbon source. By deletion and mutational analysis of sequences upstream of this gene, we have identified a region which is involved in the regulation of transcription. It is contained within a 52-base-pair sequence, UAST52 (upstream activation sequence thiolase 52), located between 203 and 151 nucleotides upstream of the translational initiation codon. This sequence proved to be required for repression, derepression and induction of transcription, and was able to activate transcription from the truncated version of the heterologous iso-1-cytochrome-c (CYC1) promoter in a similar way as in the wild-type promoter context. Sequence comparison revealed that the UAST52 contained a sequence motif ('beta-oxidation box') that is very similar to sequences located in the 5'-upstream regions of the genes coding for two other beta-oxidation enzymes of S. cerevisiae: the peroxisomal acyl-CoA oxidase and the peroxisomal trifunctional beta-oxidation enzyme of S. cerevisiae. Mutational analysis of the 'beta-oxidation box' indicates that this sequence motif acts as a UAS in vivo. Sequence comparison also revealed that just upstream of the 'beta-oxidation box', between positions -213 and -201, a potential binding site occurred for the yeast multifunctional autonomously replicating sequence binding factor ABF1. Gel-retardation-competition experiments indicate that ABF1 binds specifically to this sequence.

Acetyl-CoA C-Acyltransferase↗
PubMed1991

PAS1, a yeast gene required for peroxisome biogenesis, encodes a member of a novel family of putative ATPases.

PAS genes are required for peroxisome biogenesis in the yeast S. cerevisiae. Here we describe the cloning, sequencing, and characterization of the PAS1 gene. Its gene product, Pas1p, has been identified as a rather hydrophilic 117 kd polypeptide. The predicted Pas1p sequence contains two putative ATP-binding sites and reveals a structural relationship to three other groups of proteins associated with different biological processes such as vesicle-mediated protein transport (NSF and Sec18p), control of cell cycle (Cdc48p, VCP, and p97-ATPase), and modulation of gene expression of the human immunodeficiency virus (TBP-1). The proteins share a highly conserved domain of about 185 amino acids including a consensus sequence for ATP binding. We suggest that these proteins are members of a novel family of putative ATPases and may be descendants of one common ancestor.

Adenosine Triphosphatases↗
PubMed1991

Yeast cell cycle protein CDC48p shows full-length homology to the mammalian protein VCP and is a member of a protein family involved in secretion, peroxisome formation, and gene expression.

Yeast mutants of cell cycle gene cdc48-1 arrest as large budded cells with microtubules spreading aberrantly throughout the cytoplasm from a single spindle plaque. The gene was cloned and disruption proved it to be essential. The CDC48 sequence encodes a protein of 92 kD that has an internal duplication of 200 amino acids and includes a nucleotide binding consensus sequence. Vertebrate VCP has a 70% identity over the entire length of the protein. Yeast Sec18p and mammalian N-ethylmaleimide-sensitive fusion protein, which are involved in intracellular transport, yeast Pas1p, which is essential for peroxisome assembly, and mammalian TBP-1, which influences HIV gene expression, are 40% identical in the duplicated region. Antibodies against CDC48 recognize a yeast protein of apparently 115 kD and a mammalian protein of 100 kD. Both proteins are bound loosely to components of the microsomal fraction as described for Sec18p and N-ethylmaleimide-sensitive fusion protein. This similarity suggests that CDC48p participates in a cell cycle function related to that of N-ethylmaleimide-sensitive fusion protein/Sec18p in Golgi transport.

Adenosine Triphosphatases↗
PubMed1989

Isolation of peroxisome-deficient mutants of Saccharomyces cerevisiae.

Two mutants of Saccharomyces cerevisiae affected in peroxisomal assembly (pas mutants) have been isolated and characterized. Each strain contains a single mutation that results in (i) the inability to grow on oleic acid, (ii) accumulation of peroxisomal matrix enzymes in the cytosol, and (iii) absence of detectable peroxisomes at the ultrastructural level. These lesions (pas1-1 and pas2) are shown to be nonallelic and recessive. Crossing of pas1-1 and pas2 strains resulted in diploid cells that had regained the ability to grow on oleic acid as sole carbon source and to form peroxisomes. These pas mutants may provide useful tools for future studies on the molecular mechanisms involved in peroxisomal assembly.

Acetyl-CoA C-Acetyltransferase↗
PubMed1989

The effect of the phenylalkylamine D888 (devapamil) on force and Ca2+ current in isolated frog skeletal muscle fibres.

1. The effects of the (+)- and the (-)-isomer of the phenylalkylamine derivative D888 (desmethoxyverapamil or devapamil) on isometric force and slow Ca2+ inward current were investigated in short toe muscle fibres of the frog (Rana temporaria). The experiments were performed under voltage-clamp conditions with two flexible internal glass microelectrodes at 10 degrees C in a TEA sulphate solution containing approximately 4 mM-free Ca2+. 2. In the presence of 0.05-5 microM-(-)-D888 a normal phasic contracture could be induced by a depolarizing voltage step. When depolarization was maintained for some minutes the force-controlling system turned into a stabilized inactivated state (paralysis) from which it recovered upon repolarization within minutes instead of seconds. With the (+)-isomer (0.5-20 microM), a similarly retarded restoration was observed. However, it proved to be less effective than the (-)-isomer. 3. D888 caused a shift to more negative potentials of the S-shaped curve, which describes the voltage dependence of force restoration in the steady state (restoration time 15 min). The potential of half-maximum restoration in the absence of the drug (V = -35.8 mV) changed as follows. (-)-D888: -56 mV (0.05 microM), -69 mV (0.2 microM), -77.5 mV (0.5 microM), and -82 mV (5 microM); (+)-D888: -55.8 mV (0.5 microM), -76.5 mV (5 microM), and -85 mV (20 microM). 4. On the assumption that D888 binds only to the inactivated form of the voltage sensor of force control in the T-tubular membrane (modulated receptor hypothesis) the data presented in paragraph 3 allowed an estimation of the drug-receptor dissociation constants. The KD values ascertained in this way, 1.71 nM for the (-)-isomer and 12.9 nM for the (+)-isomer, are in fair agreement with those obtained from [3H]D888 binding studies by other authors. 5. A comparison between equal concentrations of the two isomers regarding their effect on the speed of restoration and the time needed to transform the sensor into the paralysed state suggests that the differences in the dissociation constants are mainly due to a greater dissociation rate of the (+)-isomer from the sensor. 6. The restoration of the Ca2+ channel was retarded by D888 to a similar extent as that of the voltage sensor. This parallel action on both systems indicates structural similarities between the voltage sensor and the Ca2+ channel. 7. It is concluded that D888 'stabilizes' the inactivated state of the voltage sensor and the Ca2+ channel in a way similar to D600, but with a higher potency. Both isomers of D888 showed an antagonistic action and differed only in their potency.

Animals↗
PubMed1987

[Fairy tale drama--an inpatient group psychotherapeutic method for patients with ego disorders].

Patients with "early disturbances" partly display considerable resistance to treatment methods that promote regression or aim at uncovering inhibitions, since they themselves lack emotional differentiation. In an inpatient setting such patients are able to transfer their non-experienced share of feelings and references to the treating personnel. If these transferred feelings are not properly processed and managed, therapy blockades and communication blockades arise, resulting in considerable frustrations of all concerned within the therapeutic community. Against the background of these experiences we developed a ward group method inviting both patients and staff to join hands in creative playing (e.g. by acting a fairy tale on the stage); within this framework it is possible to shape latent group, ward and team conflicts on a metaphorical level. As a result we achieve a reduction of the emotional tension level within the ward, a reduction of team conflicts, and hence an improved therapeutical climate and better possibilities to translate into reality appropriate strategies for a fair solution of problems. The overall framework, the therapeutic concept, as well as the individual stages of work-in-progress in the realisation of the fairy-tale drama are described in detail (e.g. phase of birth of the fairy tale, phase of fairy-tale recital, phase of casting, actual performance phase and subsequent discussion phase). These steps are described both as regards their form and their contents; they are compared with other group techniques of a similar nature.

Ego↗