Nature of the plasmid-linked penicillinase regulatory region in Staphylococcus aureus

Summary Four noninducible staphylococcal penicillinase plasmids reported to produce a very low basal level of penicillinase were investigated. Incorporation of 5-methyltryptophan, which is known to inactivate the operator binding site of wild-type penicillinase repressor and thereby elicit penicillinase synthesis, did not induce penicillinase synthesis in any of these micro mutants. Therefore, these plasmids are not simply peni ^S mutants. Heterodiploid strains composed of a plasmid fully constitutive for penicillinase synthesis and one of the various micro penicillinase plasmids were constructed. Three of these heterodiploids produce a normal basal level of penicillinase and are inducible by 5-methyltryptophan but not by the standard gratuitous penicillinase inducer. Therefore, each of these three noninducible micro plasmids produces a peni ^S repressor, but in addition, each must bear a mutation in the penZ region. The fourth heterodiploid produces a fully constitutive level of penicillinase. The noninducible micro plasmid present in this heterodiploid must contain a penI ^– mutation and a mutation in the penZ region. Consequently, each of these four noninducible micro plasmids contains at least two mutants genes. Hence, the phenotype of noninducibility plus low basal penicillinase is not due to a point mutation in a second penicillinase regulatory region as has been proposed. Instead, these results strongly suggest that there is only one penicillinase regulatory gene located on the penicillinase plasmid and that this gene (penI) specifies the penicillinase repressor.Journal Paper No. J-8700 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa; Project No. 2029     

Conformational energy analysis of the leucine repeat regions of C/EBP, GCN4, and the proteins of the myc, jun, and fos oncogenes

It has been recently proposed that certain DNA binding proteins (including C/EBP, GCN4 and themyc, jun, andfos oncogene proteins) share a common structural motif based on helix-promoting regions containing heptad repeat sequences of leucines. It has been suggested that this structure is critical to the biological activity of these proteins, since it facilitates the formation of functional dimers held together by interdigitating leucine side-chains along the hydrophobic interfaces between long -helical regions of the polypeptide chains in a configuration termed the leucine zipper. In this paper, conformational energy analysis is used to determine the preferred three-dimensional structures of the leucine repeat regions of these proteins. The results indicate that, in all cases, the global minimum energy conformation for these regions is an amphipathic -helix with the leucine side-chains arrayed on one side in such a way to favor leucine zipper dimerization. Furthermore, amino acid substitutions in these regions (such as Pro for Leu), that are known to inhibit dimer formation and prevent DNA binding, are found to produce significant conformational changes that disrupt the amphipathic helical structure. Thus, these results provide support for the proposed leucine zipper configuration as a critical structural feature of this class of DNA binding proteins.     

Testis-specific β2 tubulins are identical in Drosophila melanogaster and D. hydei but differ from the ubiquitous β1 tubulin

In Drosophila as in many organisms tubulins are encoded by a gene family. We have determined the complete nucleotide sequences coding for the 1 and 2 tubulins of Drosophila melanogaster and the 2 tubulin of D. hydei, and found these insect tubulins to be highly conserved and like tubulins of other organisms. This is discussed with reference to the possible functional domains of these proteins. — The 1 tubulin gene of Drosophila is constitutively expressed, whereas the 2 tubulin is expressed specifically in the testes. In D. melanogaster the amino acid sequences of these proteins are 95% homologous, differing at only 25 positions. In the testes the 2 tubulin participates in different microtubules as shown by genetic analysis (Kemphues et al. 1982). Interestingly, all of the amino acids characteristic of the testis-specific 2 tubulin are also present in the corresponding gene of D. hydei. Of special interest is the high degree of conservation of the carboxy-terminal domain in these functionally equivalent tubulins.     

Utilization of dimeric lignin model compounds by mixed bacterial cultures

Summary The degradation of dimeric phenylpropanoid lignin model compounds using mixed bacterial cultures was studied. The six model compounds contained the most common linkages of lignin: -O-4, -, -5, and -1. The results indicate that it is possible to enrich bacteria which are able to degrade all these compounds. Bacteria were also able to use these dimers as the sole source of carbon for growth. In view of these results it seems probable that bacterial inability to degrade polymeric lignin is due to the physical properties such as the molecular size of lignin.     

Theory structure,reduction, and disciplinary integration in biology

This paper examines the nature of theory structure in biology and considers the implications of those theoretical structures for theory reduction. An account of biological theories as interlevel prototypes embodying causal sequences, and related to each other by strong analogies, is presented, and examples from the neurosciences are provided to illustrate these middle-range theories. I then go on to discuss several modifications of Nagel's classical model of theory reduction, and indicate at what stages in the development of reductions these models might best apply. Finally I consider several implications of these analyses of theory structure and reduction for disciplinary integration in biology.     

Secondary H/D isotope effect on hydrogen-bonded hydroxyl groups as a tool for recognizing distance constraints in conformational analysis of oligosaccharides

An isotopomer-selected NOE (ISNOE) method for the unequivocal identification of mutually hydrogen-bond-linked hydroxyl groups is described. It relies on the fact that the OH group's signal patterns obtained for a partially deuterated sample originate from both isotopomers of the partner hydroxyl, whereas a NOE for this group can originate from cross-relaxation with the protio isotopomer of this hydroxyl only. Hence, the isotopically shifted component of this group's signal does not appear in a ROE difference spectrum obtained with selective excitation of the partner hydroxyl. This method is also applicable in those cases when only one of two mutually hydrogen-bonded groups exhibits resolvable isotope shifts. Furthermore, it is shown that isotope shifts may occur even for pairs of OH groups that are not mutually hydrogen-bonded, if these participate in hydrogen bonds with other hydroxyls and thereby affect conformational equilibria. The ISNOE experiment enables one to distinguish between these two sources of isotope shifts. Since the OO distance for hydrogen-bonded hydroxyls in sugars is known to lie between 2.7 and 3.0 Å , the hydrogen bonds established by ISNOE can be used in conformational analysis as reliable, motionally non-averaged distance constraints for the conformations containing these bonds.     

Interactions of protein kinase CK2 subunits

Several approaches have been used to study the interactions of the subunits of protein kinase CK2. The inactive mutant of CK2 that has Asp 156 mutated to Ala (CK2A156) is able to bind the CK2 subunit and to compete effectively in this binding with wild-type subunits and . The interaction between CK2A156 and CK2 was also demonstrated by transfection of epitope-tagged cDNA constructs into COS-7 cells. Immunoprecipitation of epitope-tagged CK2A156 coprecipitated the subunit and vice-versa. The assay of the CK2 activity of the extracts obtained from cells transiently transfected with these different subunits yielded some surprising results: The CK2 specific phosphorylating activity of these cells transfected with the inactive CK2A156 was considerably higher than the control cells transfected with vectors alone. Assays of the immunoprecipitated CK2A156 expressed in these cells, however, demonstrated that the mutant was indeed inactive. It can be concluded that transfection of the inactive CK2A156 affects the endogenous activity of CK2. Transfection experiments with CK2 and subunits and CK2A156 were also used to confirm the interaction of CK2 with the general CDK inhibitor p21WAF1/CIP1 co-transfected into these cells. Finally a search in the SwissProt databank for proteins with properties similar to those derived from the amino acid composition of CK2 indicated that CK2 is related to protein phosphatase 2A and to other phosphatases as well as to a subunit of some ion-transport ATPases.     

Saponin-like substances inhibit α-galactosidase production in the endosperm of fenugreek seeds

When fenugreek (Trigonella foenum-graecum L.) endosperms plus testa (endosperms), which had been isolated from 5-h-imbibed seeds, were incubated for at least 2 h under germination conditions, they leaked substances which, like exogenous abscisic acid (ABA), inhibited the production of fenugreek endosperm -galactosidase. However, unlike ABA, 8 h treatment with these inhibitors had no effect on fenugreek endosperms which had been isolated from 15-h-imbibed seeds and leached for 2 h. This indicated that either their inhibitory action was on processes which were related to the production of -galactosidase and had been completed by this time, or that there might be factors present which inactivate these inhibitors. It was also concluded that the action of the endosperm leachate could not be attributed to the presence of ABA. The activity of the leachate decreased when it originated from endosperms imbibed for periods longer than 25 h and thin-layer chromatography (TLC) of extracts from these endosperms showed decreased contents of the leachable inhibitors as imbibition proceeded. From the seed leachate, which had a TLC pattern and inhibitory action similar to that of the endosperm, were isolated three substances which, when applied to endosperms, inhibited the production of -galactosidase activity. According to their chromatographic behaviour and their reaction with specific reagents, there are strong indications that these substances are saponins. These diffusible saponin-like substances were located in both endosperm and perisperm and their physiological role is discussed.Abbreviations ABA abscisic acid - PEG polyethylenglycol - TLC thin-layer chromatography We wish to thank the Alexander S. Onasis Public Benefit Foundation for a grant to K.Z. and Dr. J.S.G. Reid (University of Stirling, Scotland) for a kind gift of fenugreek seeds.     

Deep tectal cells in pigeons respond to kinematograms

Summary Deep tectal neurons in pigeons respond selectively to moving visual stimuli, and are inhibited by large background patterns moved in-phase with these stimuli. In this investigation we demonstrate that these same deep tectal neurons respond equally well to kinematograms as they do to traditional luminance contrast stimuli typically employed in visual experiments.Computer generated kinematograms, the motion domain equivalents of random dot stereograms, were used as stimuli in these experiments. These kinematograms, where a small centrally located set of random dots is moved coherently in one direction while the remaining dots are moved in a different direction, thus constitute a pure motion stimulus where the stimulus form is only visible in the dynamic pattern, but does not exist on any single frame. Both object configured and hole configured kinematograms were employed; the former appearing as regions of texture moving over, or in front of, the background texture, while the latter appear as windows through which a more distant textured surface is revealed.Extracellular recordings from isolated deep tectal cells showed that all units responded in a very similar manner whether the stimulus was an object configured kinematogram or the more traditional luminance contrast variety. This similarity included directional selectivity, the in-phase inhibition anti-phase facilitation effect, and sensitivity to opposed motion independent of direction. However, when the kinematograms were configured as holes none of the units tested responded to these stimuli. The significance of these observations for tectal functioning, image segmentation through motion and animal camouflage is discussed.     

Calcium-Dependent Regulation of Interactions of Caldesmon with Calcium-Binding Proteins Found in Growth Cones of Chick Forebrain Neurons

1. This study was undertaken to determine if caldesmon, calmodulin, S100, and neurocalcin were present in chick forebrain neurons, and if so, to investigate the interactions of these proteins in the presence of different concentrations of calcium.2. Immunocytochemistry was used to determine the presence and localization of these proteins in cultured forebrain neurons. Western blotting, gel electrophoresis in the presence of different concentrations of calcium, chemical cross-linking, and affinity chromatography were used to investigate the interactions of these proteins with each other.3. Our data show that caldesmon and three calcium-binding proteins (S100, calmodulin, and neurocalcin ) are localized in growth cones and neurites of chick forebrain neurons in culture. In the presence of different concentration of calcium, these calcium-binding proteins have different affinities to caldesmon and to each other. S100 binds with greater affinity than calmodulin to caldesmon, and its ability to bind to caldesmon is regulated by neurocalcin .4. These findings suggest a specific calcium-dependent regulatory pathway for modulating actomyosin during growth cone motility.     

Migration and life histories of the threespine stickleback: evidence for adaptive variation in growth rate between populations

Synopsis Migratory fishes should exhibit higher growth rates and larger body size than nonmigrants. To test this hypothesis, laboratory reared threespine sticklebacks,Gasterosteus aculeatus, representing three populations from a single drainage in northern California, U.S.A. were compared. These populations differ in their migratory patterns, ranging from highly migratory anadromous forms to nonmigratory freshwater residents. The nonmigratory inland population was significantly smaller in standard length at most ages, with correspondingly reduced growth rates compared to the more migratory upstream and estuary populations. Von Bertalanffy growth functions described the growth trajectories of these fish well, accounting for approximately 99% of the variance in body size exhibited by these populations. Broad sense heritability estimates for body size were significant for all three populations at age 60 days but were significant only for the inland population at later ages. The results of this study provide evidence of genetically-based variation in growth rate and body size among these stickleback populations, and these differences are consistent with the hypothesis that adaptation to different migratory lifestyles has occurred.     

Considerations on the astroglial architecture and the columnar organization of the cerebral cortex

Present knowledge on astroglial roles in brain organization and function indicates that these cells can regulate the extracellular ionic composition and modulate neuronal activity. In this regard, the panglial synctium formed by electrically and chemically coupled stellate astrocytes (general mammalian architecture) is believed to provide an intracellular pathway for the redistribution of ions and molecules within the cerebral cortex. Long astroglial interlaminar processes (primate-specific architecture) that run parallel to apical dendrites in the cerebral cortex of primates, may provide additional properties to the glial participation in cortical physiology and function. Since these processes are exclusively present within the primate order, functional models of cortical computations for these species should incorporate the astroglial interlaminar architecture in addition to the panglial synctium. This study analyzes possible implications of interlaminar astroglial processes, for the regulation of the extracellular ionic composition and segregation of functional columns in the cerebral cortex.     

Adrenergic and muscarinic receptor regulation and therapeutic implications in heart failure

In end-stage heart failure the expression of different myocardial regulatory proteins involved in the -adrenergic cAMP signalling pathway is altered. The downregulation of -adrenoceptors and their uncoupling from the effector as well as an increased expression of the inhibitory GTP-binding protein seem to be the most important alterations. Since catecholamine levels are elevated in these patients and since some alterations can be restored after treatment with -adrenoceptor antagonists it was hypothesized that excessive -adrenergic stimulation could be involved in these alterations.In this article the changes of -adrenergic receptors, GTP-binding proteins, sarcoplasmic reticulum Ca²⁺-ATPase and of phospholamban found in heart failure are addressed with its possible therapeutic implications.     

Ritual,the state,and the transformation of emotional discourse in Iranian society

This paper explores the social and cultural organization of Iranian emotional discourse and its transformation in post-revolutionary Iran. First, the Moharram dramas we participated in during field research are described, indicating how these performances organized a prototypical view of the social order, the self, and the passions. Using Kapferer's distinction between transcendental and transformative rituals, we argue that these dramas were traditionally organized as transcendental rites. Second, data on grieving rituals and depressive illness among Iranians is introduced, focusing on the transformative qualities of mourning rites and suggesting an interpretation of depression as a failure of the work of culture. Third, the appropriation of these symbolic forms of society, self, and the emotions by the current Iranian Islamic state and the role of the state in defming the meaning and legitimacy of emotions and their expression is analyzed.     

How natural is a nature reserve?: an ideological study of British nature conservation landscapes

Areas set apart for nature conservation in Britain are broadly categorised according to their cultural purpose, and names are assigned to these in this paper. Nature reserves may be similar to zoos and botanic gardens in aiming to maintain the diversity of species and if so are termed biodiversity reserves. This tradition understands nature as a static collection of entities apart from humans. Maintaining traditional management at a site is arguably a good way to sustain species, it also retains old ways in which humans and nature were integrated in the life of the nation and so are called historic countryside parks. There is growing interest in wilderness areas, where nature is seen as primarily processes protected from human interference. Despite the strength of each of these, they suffer from attempting to restrict nature to a ghetto, a process that is economically and environmentally costly. Companion places are places which set sustainable examples of integrating human life and economic activity with maintaining biodiversity and offering an opportunity to encounter wild processes at the heart of life. The language of these four types, or vectors, of nature reserves is offered to help the discussion of our place in nature.     

The α and γ subunits of 7S nerve growth factor are present in excess concentrations in the mouse submandibular gland

Summary The 7S nerve growth factor molecule, found in the mouse submandibular gland, is comprised of three distinct protein subunits named , and -NGF. In this paper, radioimmunoassays specific for each subunit were used to measure the concentrations of these subunits in homogenates of mouse submandibular gland. It was determined that there were excess concentrations of both the and subunits, more than enough to bind all of the -NGF in the gland to form 7S-NGF. The radioimmunoassay data was confirmed by gel filtration experiments. In the gel filtration experiments, the excess and subunits eluted at positions which would indicate that these excess subunits were free and not bound in the 7S-NGF complex. The identity of the excess and subunits was substantiated by ion exchange chromatography, isoelectric focusing polyacrylamide gels and immunoblotting experiments. In conclusion, there are considerable quantities of and subunits present in the submandibular gland which are not bound to -NGE The functional significance of these excess concentrations of the and subunits is not known.     

Synonymous nucleotide substitution rates of β-tubulin and histone genes conform to high overall genomic rates in rodents but not in sea urchins

Summary Sea urchin and rodent genomes have been posited to evolve rapidly as indicated by divergences in single copy nuclear DNA sequences. We have examined whether the synonymous substitution rates of three highly conserved genes, -tubulin, histone H4, and histone H3, adhere to these high genomic substitution rates by comparing sequences between two sea urchins,Strongylocentrotus purpuratus andLytechinus pictus, and between rodents and humans. Whereas the rate of change between the 3 untranslated regions of the -tubulin cDNA ofS. purpuratus (Sp-1), sequenced in this study, and ofL. pictus (Lp-3) was consistent with the overall rate of change estimated from previous DNA hybridization results between these species, the synonymous substitution rates for the carboxyl domains of these -tubulins, as well as for the late histones H4 and H3, were significantly depressed. In contrast, synonymous nucleotide substitution rates between rodents and between rodent and human for the carboxyl domain proper of identical -tubulin isotypes and for histone H4 and H3.1 did not differ from the overall rate of change for the rodent genomes. Moreover, an analysis of paralogous human and mouse -tubulin sequences supported the conclusion that the synonymous substitution rates in the mouse were higher than those in the human. Differences in constraint on evolutionary change were not evident strictly from the conserved amino acid sequences and base compositions of these genes. Other constraining influences seemed more relevant to the departure of the synonymous substitution rates of the sea urchin -tubulin and histone coding regions from the average genomic rate.     

Restriction fragment length polymorphisms as genetic markers in soybean,Glycine max (L.) merrill

Summary Restriction Fragment Length Polymorphisms (RFLP) have been identified between widely distant cultivars (Minsoy and Noir 1 ) of soybean Glycine max (L.) Merrill. Using as probes randomly chosen clones of DNA, one in five probes revealed a polymorphism. More than half of these polymorphisms appear to result from rearrangements of the genomic DNA. Twenty seven markers were analyzed for linkage in F₂ plants. Eleven of these markers were contained in four linkage groups. Five cultivars were compared in a search for new alleles. When RFLP markers corresponding to low copy DNA were used to analyze three other cultivars — Sooty, Forrest and Mandarin (Ottawa) — few new alleles were found. Using these probes, five different markers could be used to differentiate the five cultivars. Complex probes, which correspond to repeated DNA, revealed different polymorphisms in different cultivars and a single such probe could be used to distinguish the five cultivars from each other.     

A search for mutations that affect the non-oxidative pentose phosphate pathway inSaccharomyces cerevisiae

A number of selection strategies have been devised to obtain mutations in the non-oxidative pentose phosphate pathway in the yeastSaccharomyces cerevisiae. Some of these schemes exploit the bidirectionality of this pathway by selecting for mutants that can grow on a mixture of two carbon sources, which allow the pathway to function in opposite directions, while failing to utilize either of them alone. Other strategies select for mutations that prevent growth on glucono-5-lactone or xylulose, compounds metabolized exclusively through the pentose phosphate pathway. In the present study, these schemes yielded 35 mutants that define 11 genetic complementation groups. None of these mutations, however, affected the activity of the pathway enzymes, assayed in cell-free extracts. Nevertheless, the mutants were deficient in various aspects of carbohydrate metabolism and in the biosynthesis of the aromatic amino acids. The problems associated with the selection of mutants directly affected in the functioning of the enzymes of the non-oxidative pentose phosphate pathway have been discussed.     

Somaclonal variation in wheat: genetic and cytogenetic characterisation of alcohol dehydrogenase 1 mutants

Summary The progeny of 551 regenerants of the hexaploid wheat cultivar Millewa were analysed for somaclonal mutants at the threeAdh-1 loci in hexaploid wheat. Seventeen regenerants gave rise to progeny having altered ADH1 zymograms. Progeny with altered zymograms in 13 of these regenerants were aneuploid. The remaining 4 regenerants gave rise to euploid progeny with altered ADH1 zymograms. The genetics of three of these somaclonal mutants is described in detail. These regenerants were interpreted to possess a 4A isochromosome, a 3BS/4A translocation and a 7BS/4A translocation, respectively.This research was partly supported by the Rockefeller Foundation and a Reserve Bank of Australia Rural Credit Development Fund research grant