Identification and characterization of a defective SSV1 genome integrated into a tRNA gene in the archaebacterium Sulfolobus sp. B12

Summary Within the chromosome of the archaebacterium Sulfolobus sp. B12, a 7.4 kb region was identified which displayed extensive sequence similarities to the 15.5 kb genetic element SSV1 carried by the same strain both as a circular form and as a site-specifically integrated copy. DNA sequence analysis indicated that this 7.4 kb region (designated SSV1^intB) represented an SSV1-like element distinguishable from the full-length integrated copy (designated SSV1^intA) by extensive deletions and point mutations. The physical organization of DNA sequences of SSV1^intB indicated that this element was integrated at the same attP site as previously identified for SSV1^intA. A comparison of the DNA sequences at the left attachment sites of SSV1^intA and SSV1^intB revealed that they both represented very similar putative arginine tRNA genes followed by a 10 by inverted repeat sequence. S1 nuclease mapping experiments indicated that these tRNA genes are transcribed. Offprint requests to: W. Zillig     

Misting and nitrogen fertilization of shoots of a saltmarsh grass: effects upon fungal decay of leaf blades

We conducted a 12-week field manipulation experiment in which we raised the nitrogen availability (ammonium sulfate fertilization to roots) and/or water potential (freshwater misting) of decaying leaf blades of a saltmarsh grass (smooth cordgrass, Spartina alterniflora) in triplicate 11-m² plots, and compared the manipulated plots to unmanipulated, control plots. The ascomycetous fungi that dominate cordgrass leaf decomposition processes under natural conditions exhibited a boosting (>2-fold) of living standing crop (ergosterol content) by misting at the 1 st week after tagging of senescent leaves, but afterwards, living-fungal standing crop on misted blades was equivalent to that on control blades, confirming prior evidence that Spartina fungi are well adapted to natural, irregular wetting. Misting also caused 2-fold sharper temporal declines than control in instantaneous rates of fungal production (ergosterol synthesis), 5-fold declines in density of sexual reproductive structures that were not shown by controls, and 2-fold higher rates of loss of plant organic mass. Extra nitrogen gave a long-term boost to living-fungal standing crop (about 2-fold at 12 weeks), which was also reflected in rates of fungal production at 4 weeks, suggesting that saltmarsh fungal production is nitrogen-limited. Although bacterial and green-microalgal crops were boosted by manipulations of nitrogen and/or water, their maximal crops remained 0.3 or 2% (bacteria or green microalgae, respectively) of contemporaneous living-fungal crop. The fungal carbon-productivity values obtained, in conjunction with rates of loss of plant carbon, hinted that fungal yield can be high (>50%), and that it is boosted by high availability of nitrogen. We speculate that one partial cause of high fungal yield could be subsidy of fungal growth by dissolved organic carbon from outside decomposing leaves.     

Identification of five hemoglobins in B6C3F1 mice by mass spectrometry and sequence analysis

The aim of the work is to identify and characterize the hemoglobins found in B6C3F1 mice using mass spectrometry. The primary structures are compared to those reported for BALB/c mice. Individual hemoglobin chains were isolated by reversed-phase high performance liquid chromatography (RP-HPLC). The molecular masses of the globins were determined using electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI). The purified globin chains were enzymatically cleaved and the resulting peptides were separated by RP-HPLC. The chains were identified by N-terminal sequencing and mass spectrometry (MALDI). Selected peptides were analysed by Edman degradation. ESI analysis indicates that B6C3F1 mice have two -globin chains (-1 and -2) and at least three β-globin chains, β-1, β-2 and β-3. This is one additional - and one additional β-globin chain than reported in the literature for BALB/c mice. Mass and sequence analysis of enzymatically generated peptides showed variations in the amino acid sequence in the -1, -2, β-2 and β-3 chains compared to the BALB/c mouse hemoglobins (, β^minor and β^major). The study showed that mass spectrometry in combination with traditional protein chemistry is able to identify and locate minor protein sequence variations.     

Ammonia volatilization from tropical legume mulches and green manures on unlimed and limed soils

In the humid tropics, legumes are harvested and surface applied as mulch or incorporated as green manure. Studies on N dynamics and budgets from these systems report unaccounted losses of N. Ammonia volatilization may account for a significant percentage of these unexplained N deficits. The main objectives of this study were to: 1) determine the rate and amount of ammonia volatilization from organic amendments, both incorporated (green manure) and unincorporated (mulch), 2) compare ammonia volatilization of organic amendments on both acid (unlimed) and limed soils, and 3) correlate quality, i.e. polyphenolic and lignin concentration and carbon-to-nitrogen ratio, of the organic amendments with ammonia volatilization and net N mineralization. In an incubation experiment, ammonia volatilization losses and net N mineralization were measured from fresh leaflets of 10 legumes over a three-week period. Ammonia volatilization losses for the 10 species ranged from 3.4 to 11.8% of the total N applied in the organic amendment. Lignin content was negatively correlated to ammonia volatilization. Ammonia volatilized from mulches but not green manures, on both unlimed and limed soils, suggesting that ammonia volatilization is a surface phenomenon and not affected by soil pH. Net N mineralization was affected by species and soil pH, but was unaffected by placement (green manure or mulch). For the farmer in low-input agriculture where N tends to be limiting, volatilization losses of N from legume mulch systems could be on the same order of magnitude as crop removal.     

Evolutionary stable strategies and game dynamics for n-person games

This note contains a generalization of the definition of an evolutionary stable strategy and of the corresponding game dynamics from 2-person to n-person games. This broader framework also allows modelling of several interacting populations or of populations containing different types of individuals, for example males and females.     

On representation and approximation of nonlinear systems

Many methods for the analysis of nonlinear systems rely on a Volterra system-representation in terms of integral kernels. This paper considers two questions: 1) whether Volterra-like representations are possible for all smooth systems (i.e. analytic operators),-2) which classes of systems can be approximated by some interesting subclasses of the smooth systems, e.g. Volterra systems with separable kernels and sandwich systems (cascades of linear systems and nomemory nonlinearities). Whereas the answer to question 1 is positive, the answer to question 2 depends on the type of approximation, i.e. topology, that is used.     

Rabbit muscle phosphorylase derivatives with oligosaccharides covalently bound to the glycogen storage site

Linear maltooligosaccharides, e.g., maltoheptaose or terminal 4-O-methylmaltoheptaose, activated by cyanogen bromide, react covalently with rabbit muscle phosphorylases b and a (EC 2.4.1.1). Site-specific modification prevents further binding to glycogen and shifts the phosphorylase a tetramer-dimer equilibrium in favor of the dimer. Use was made of these properties to separate by affinity chromatography and gel filtration phosphorylase a dimers with specifically bound oligosaccharide from unspecifically modified products. The phosphorylase a-maltoheptaose derivative carries one oligosaccharide residue per monomer and can be distinguished from the native enzyme by its electrophoretic mobility in polyacrylamide gels or by affinity electrophoresis. Phosphorylase a preparations with covalently bound maltooligosaccharides are enzymatically active in the presence of a primer and alpha-D-glucopyranose 1-phosphate (glucose-1-P). Methylation of the nonreducing chain terminus of the bound oligosaccharide has no effect on glycogen synthesis. These findings exclude the participation of bound oligosaccharides in chain elongation. Purified covalent phosphorylase a-maltoheptaose complexes are stable dimers. They are no longer activated by glycogen. The properties of covalently modified phosphorylase-oligosaccharides are consistent with and provide direct evidence for the existence of a glycogen storage site in rabbit muscle phosphorylases. Covalent occupation of the storage site renders the affinity of glucose-1-P to phosphorylase a independent of modulation by glycogen, supporting the assumption that the glycogen storage site is involved in interactions with the catalytic site.     

Identification of a Gs-protein coupling domain to the beta-adrenoceptor using site-specific synthetic peptides. Carboxyl terminus of Gs alpha is involved in coupling to beta-adrenoceptors

Competition between Gs-protein and the synthetic peptide, GSA 379-394, derived from the carboxyl-terminal region of the alpha s-subunit, led to complete inhibition of receptor-mediated adenylate cyclase activation in turkey erythrocyte membranes. Related peptides corresponding to the homologous carboxyl-terminal region of alpha t-, alpha il- or alpha o-subunits did not interfere with beta-receptor-Gs coupling. The direct coupling between Gs and adenylate cyclase was not influenced by any of these peptides. These results emphasize the important role of the carboxyl-terminus of G-protein alpha-subunits for the specific recognition of their corresponding receptors and for signal transduction.     

Identification of a mutation within the structural gene for the α subunit of DNA-dependent RNA polymerase of E. Coli

Summary An E. coli mutant rpoA109 unable to support the growth of phage P2 produces DNA-dependent RNA polymerase with an altered subunit. Histidine is substituted for leucine in one tryptic peptide from the mutant subunit. The existence of only one rpoA gene within the E. coli chromosome is indicated.     

Letter: Preliminary x-ray data from well-ordered crystals of a human immunoglobulin G molecule

Crystals of an intact immunoglobulin G molecule, which are suitable for high resolution X-ray analysis, have been grown. The space group is P3₁21 (or enantiomorph) and the crystallographic diad relates the two halves of one molecule.