Studies on an interesting Saccharomyces carlsbergensis mutant

Summary 32 sputa from 29 patients known to have yeasts were emulsified and quantitatively plated on Sabouraud's glucose agar. Colony counts showed that the yeast flora varied from about 300 to about one million cells per ml of the specimen. Sputa collected at bed-side and plated within 2 hours gave lower counts than those received routinely. In the 28 specimens in which identification was carried outC. albicans was found in 23 as the only or the dominant species andC. tropicalis in 4; an unidentified species was present in one specimen.This work was supported by a Grant from the Medical Research Council, Ottawa, Canada.     

Studies on an interesting Saccharomyces carlsbergensis mutant

The genetics of a spontaneous double mutant Hla, belonging toSaccharomyces carlsbergensis, has been studied. The two mutated loci, controlling the biosyntheses of lysine and uracil are either loosely linked or they are located on separate chromosomes. Spontaneous mutations in the two genes are therefore likely to have followed from two independent mutagenic events. The mating-type locus, the methionine, the lysine and the uracil synthesising loci are not linked. Segregations for mating-type locus are Mendelian where as segregations for lysine, uracil and methionine synthesising loci are both Mendelian and Non-Mendelian. The 4:0 class of Non-Mendelian segregation is not found. The apparently Non-Mendelian segregations for mehtionine synthesis can be explained on the basis of the presence of two complementary methionine synthesising genes in Hla.

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Zusammenfassung Die Genetik eines spontanen Doppelmutanten Hla, der zuSaccharomyces carlsbergensis gehört, wurde untersucht. Die zwei Loci, die die Biosynthese von Lysin und Uracil kontrollieren, sind entweder locker gebunden oder sind an verschiedene Chromosomen gebunden. Spontane Mutationen in beiden Genen sind deshalb wahrscheinlich das Ergebnis von zwei unabhängigen mutagenen Faktoren. Der Locus der Geschlechtstype, die Loci der Synthese von Methionine, Lysine und Uracil sind nicht gebunden. Die Absonderung des Locus für die Geschlechtstype ist gemäß Mendel, während die Absonderung für Lysine, Uracil und Methionine teils gemäß, teils nicht gemäß Mendel erfolgt. Die Klasse 4 : 0 der Absonderung nicht gemäß Mendel ist nicht gefunden worden. Die scheinbare Absonderung nicht gemäß Mendel für die Synthese von Methionine kann auf Grund der Gegenwart von zwei komplementären Genen in Hla für die Synthese von Methionine erklärt werden.

     

Remark on an interesting problem in topological biology

The possibility of several homotopic classes of mappings of the graph of an organism onto the primordial graph (Bull. Math. Biophysics,16, 317–48, 1954) is considered. An application of this possibility is suggested for the theoretical determination as to what type of new biological functions may be acquired by certain cells which originally perform a different biological function.     

Ferricrocin - an ectomycorrhizal siderophore of Cenococcum geophilum

The ectomycorrhizal fungus Cenococcum geophilum was grown in low-iron medium and the excreted siderophores were extracted, purified and analyzed by HPLC. The principal hydroxamate siderophore produced, was identified as ferricrocin as confirmed by analytical HPLC, FAB-mass spectrometry and ¹H- and ¹³C-NMR spectra. Although the occurrence of ferricrocin has been shown earlier to occur in the ericoid mycorrhizal fungi, this is the first report of ferricrocin in a true ectomycorrhizal fungus which is taxonomically related to the ascomycetes.     

Cytoplasmic sulfurtransferases in the purple sulfur bacterium Allochromatium vinosum: evidence for sulfur transfer from DsrEFH to DsrC

While the importance of sulfur transfer reactions is well established for a number of biosynthetic pathways, evidence has only started to emerge that sulfurtransferases may also be major players in sulfur-based microbial energy metabolism. Among the first organisms studied in this regard is the phototrophic purple sulfur bacterium Allochromatium vinosum. During the oxidation of reduced sulfur species to sulfate this Gammaproteobacterium accumulates sulfur globules. Low molecular weight organic persulfides have been proposed as carrier molecules transferring sulfur from the periplasmic sulfur globules into the cytoplasm where it is further oxidized via the "Dsr" (dissimilatory sulfite reductase) proteins. We have suggested earlier that the heterohexameric protein DsrEFH is the direct or indirect acceptor for persulfidic sulfur imported into the cytoplasm. This proposal originated from the structural similarity of DsrEFH with the established sulfurtransferase TusBCD from E. coli. As part of a system for tRNA modification TusBCD transfers sulfur to TusE, a homolog of another crucial component of the A. vinosum Dsr system, namely DsrC. Here we show that neither DsrEFH nor DsrC have the ability to mobilize sulfane sulfur directly from low molecular weight thiols like thiosulfate or glutathione persulfide. However, we demonstrate that DsrEFH binds sulfur specifically to the conserved cysteine residue DsrE-Cys78 in vitro. Sulfur atoms bound to cysteines in DsrH and DsrF were not detected. DsrC was exclusively persulfurated at DsrC-Cys111 in the penultimate position of the protein. Most importantly, we show that persulfurated DsrEFH indeed serves as an effective sulfur donor for DsrC in vitro. The active site cysteines Cys78 of DsrE and Cys20 of DsrH furthermore proved to be essential for sulfur oxidation in vivo supporting the notion that DsrEFH and DsrC are part of a sulfur relay system that transfers sulfur from a persulfurated carrier molecule to the dissimilatory sulfite reductase DsrAB.     

Seasonal variation of water chemistry and sulfur budget in an acid-sensitive river along the Sea of Japan

A long-term declining trend of pH values has been observed for several rivers in areas of central Japan that are geologically dominated by acidic rocks such as granite, rhyolite, or chert. We monitored the seasonal variation in water chemistry in one of these rivers: the Araya River in Niigata Prefecture. During the 4-year survey period, we observed temporary acidification during the rainy and snowmelt seasons when the river flow rate abruptly increased. In the rainy season, the decrease in pH may be attributable to the dilution of almost all ions and increased leaching of NO₃ ^? from the catchment. In the snowmelt season, decreases in pH were found to be associated with peaks of SO₄ ^2?, where the SO₄ ^2? was possibly derived from that accumulated in the snowpack. The data pertaining to the river showed that it had a sensitive response to meteorological events and was likely to be acid sensitive. The estimated mass balance of the river showed that the SO₄ ^2? output exceeded the corresponding input to the Araya River catchment. Mobilization of internal sulfur accumulated in forest ecosystems might have contributed to the observed long-term acidification of this acid-sensitive river.     

Biosynthesis of the pyoverdine siderophore of Pseudomonas aeruginosa involves precursors with a myristic or a myristoleic acid chain

Pyoverdine I (PVDI) is the major siderophore produced by Pseudomonas aeruginosa to import iron. Biosynthesis of this chelator involves non-ribosomal peptide synthetases and other enzymes. PvdQ is a periplasmic enzyme from the NTN hydrolase family and is involved in the final steps of PVDI biosynthesis. A pvdQ mutant produces two non-fluorescent PVDI precursors with a higher molecular mass than PVDI. In the present study, we describe the use of mass spectrometry to determine the structure of these PVDI precursors and show that they both contain a unformed chromophore like ferribactin, and either a myristic or myristoleic chain that must be removed before PVDI is secreted into the extracellular medium.     

Biosynthesis of the iron-molybdenum cofactor and the molybdenum cofactor in Klebsiella pneumoniae: effect of sulfur source.

NifQ- and Mol- mutants of Klebsiella pneumoniae show an elevated molybdenum requirement for nitrogen fixation. Substitution of cystine for sulfate as the sulfur source in the medium reduced the molybdenum requirement of these mutants to levels required by the wild type. Cystine also increased the intracellular molybdenum accumulation of NifQ- and Mol- mutants. Cystine did not affect the molybdenum requirement or accumulation in wild-type K. pneumoniae. Sulfate transport and metabolism in K. pneumoniae were repressed by cystine. However, the effect of cystine on the molybdenum requirement could not be explained by an interaction between sulfate and molybdate at the transport level. Cystine increased the molybdenum requirement of Mol- mutants for nitrate reductase activity by at least 100-fold. Cystine had the same effect on the molybdenum requirement for nitrate reductase activity in Escherichia coli ChlD- mutants. This shows that cystine does not have a generalized effect on molybdenum metabolism. Millimolar concentrations of molybdate inhibited nitrogenase and nitrate reductase derepression with sulfate as the sulfur source, but not with cystine. The inhibition was the result of a specific antagonism of sulfate metabolism by molybdate. The effects of nifQ and mol mutations on nitrogenase could be suppressed either by the addition of cystine or by high concentrations of molybdate. This suggests that a sulfur donor and molybdenum interact at an early step in the biosynthesis of the iron-molybdenum cofactor. This interaction might occur nonenzymatically when the levels of the reactants are high.     

Chlorosomes of green sulfur bacteria: Pigment composition and energy transfer

The pigment composition and energy transfer pathways in isolated chlorosomes ofChlorobium phaeovibrioides andChlorobium vibrioforme were studied by means of high performance liquid chromatography (HPLC) and picosecond absorbance difference spectroscopy. Analysis of pigment extracts of the chlorosomes revealed that they contain small amounts of bacteriochlorophyll (BChl)a esterified with phytol, whereas the BChlsc, d ande are predominantly esterified with farnesol. The chlorosomal BChla content inC. phaeovibrioides andC. vibrioforme was found to be 1.5% and 0.9%, respectively. The time resolved absorbance difference spectra showed a bleaching shifted to longer wavelengths as compared to the Q_y absorption maxima and in chlorosomes ofC. vibrioforme also an absorbance increase at shorter wavelengths was observed. These spectral features were ascribed to excitation of oligomers of BChle and BChlc/d, respectively. One-color and two-color pump-probe kinetics ofC. phaeovibrioides showed rapid energy transfer to long-wavelength absorbing BChle oligomers, followed by trapping of excitations by BChla with a time constant of about 60 ps. Time resolved anisotropy measurements inC. vibrioforme showed randomization of excitations among BChla molecules with a time constant of about 20 ps, indicating that BChla in the baseplate is organized in clusters. One-color and two-color pump-probe measurements inC. vibrioforme showed rapid energy transfer from short-wavelength to long-wavelength absorbing oligomers with a time constant of about 11 ps. Trapping of excitations by BChla in this species could not be resolved unambiguously due to annihilation processes in the BChla clusters, but may occur with time constants of 15, 70 and 200 ps.     

Biosynthesis of the modified nucleoside Q in transfer RNA.

During biosynthesis of the modified nucleoside Q, 7-(4,5-DIHYDROXYL-1-1-CYCLOPENTEN-3-YL-AMINOMETHYL)-7-DEAZAGUANOSINE, IN TRNA, the carbon atom at position 8 in precursor molecule guanine was expelled together with the nitrogen atom N-7 in a fashion similar to that in the biosynthesis of the nucleoside antibiotic toyocamycin.