Resonance Raman characterization of the di-heme protein cytochrome c4 from Pseudomonas stutzeri

 Di-heme Pseudomonas stutzeri cytochrome c ₄ has been characterized by electronic absorption and resonance Raman spectroscopies in the ferric and ferrous forms at pH 7.5 and at room temperature. The data indicate that the two hemes are inequivalent. It is proposed that the N-terminal contains a more relaxed heme as a consequence of the relative orientation of the methionine and histidine ligands with respect to the N-Fe-N directions of the heme plane. This causes a weakening of the Fe-S bond with concomitant partial dissociation of the methionine and the formation of an Fe-aquo bond. Heme group relaxation is further accompanied by less distortion of the heme group than that associated with cytochrome c, expansion of the "core" and a negative shift of the redox potential. Received: 17 December 1996 / Accepted: 6 March 1997     

Short-term variations in catches of the pea moth,Cydia nigricana, in interacting pheromone traps

Short-term variations in the relative catch in each of two or three interacting pheromone traps for the pea moth,Cydia nigricana (F.), were investigated for traps aligned along the wind. The proportional catch in each trap varied widely, although the mean values accorded with previous estimates. Over consecutive short intervals during a single trapping period the proportion caught in the centre trap of a three-trap line was constant. The proportion caught in the upwind trap of two-and three-trap lines showed trends in time. These trends differed between trapping periods, but two lines of traps operated simultaneously gave similar results to each other. It is suggested that these results, which are predicted by a model based on various components of moth orientation behaviour, are caused by changes in systematic behavioural processes, not random effects. Possible mechanisms are discussed.

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Variations à court terme des captures deCydia nigricana dans des pièges à phéromones en interaction

Résumé Des données antérieures concernant les interactions entre des pièges à phéromone alignés le long du vent ont été utilisées pour déduire les caractéristiques du comportement d'orientation deC. nigricana. Ces données ont été introduites dans un modèle de simulation quantitative qui prédisait que quand les captures totales sur une ligne ont été regroupées sur une période globale de piégage, la proportion capturée dans chaque piège devrait aussi avoir des valeurs moyennes semblables, mais varie plus largement que précédemment indiqué. Les simulations concernant des intervalles consécutifs beaucoup plus courts pendant la même période de piégage ont suggéré une forme spécifique de cette variation.Cette note signale des variations à court terme dans la proportion capturée au piège qui confirment ces prédictions. Nous montrons que sur des intervalles consécutifs brefs pendant une simple période de piégage, la proportion capturée dans le piège central d'une ligne de 3 pièges est contstante, bien que sa valeur change suivant les périodes de piégage. La proportion capturée dans le piège face au vent de 2 ou 3 lignes de pièges suit cette tendance dans le temps, ce qui est généralement bien représenté avec des courbes simples. Ces tendances changent suivant les périodes de piégage, mais des lignes de pièges fonctionnant simultanément fournissaient des résultats similaires. On suggère que ces résultats sont dus à des changements dans des processus comportementaux systématiques et non à des effects aléatoires. Les mécanismes possibles sont discutés.

     

Characterization of the mutant lytic state in lambda expression systems

The two propagative phases of bacteriophage lambda, lysogeny and lysis, can be used in concert to enhance productivity of recombinant expression systems. Lambda vectors carrying mutations to prevent both cell lysis and lambda DNA packaging in the lytic state have been shown to yield 100% stability of the product gene in lysogeny and to produce up to 15% of total cell protein as product beta-galactosidase in a mutant lytic state.(14) Despite these mutations, partial lysis of the culture was observed following induction of the cells from a lysogenic phase into the lytic state. To understand better the phage-host cell interactions and to investigate the possible cause(s) of lysis in these highly productive expression systems, we have made a detailed study of the suppressor-free system JM105(NM1070). We have found high levels of product (15% of total cell protein as beta-glactosidase) to be due chiefly to a high-copy number of lambda DNA in the mutant lytic state. There is partial lysis of the culture even in this suppressor-free system caused by a low-level natural suppression of the amber mutation in gene S of NM1070, resulting in accumulation of lambda endolysin. We have also monitored changes in cell growth and morphology upon induction of the lysogen. There is a slight increase in cell number that follows a linear relationship with time and a 25-fold increase in cell volume during recombinat protein production in the mutant lytic state.     

Biogeographic comparisons of marine algal polyphenolics: evidence against a latitudinal trend

Summary Marine allelochemicals generally are present in greater quantity and diversity in tropical than in temperate regions. Marine algal polyphenolics have been reported as an apparent exception to this biogeographic trend, with literature values for phenolic concentrations significantly higher in temperate than in tropical brown algae. In contrast, our results, the first reported for Caribbean brown algae (orders Dictyotales and Fucales), show that many species have high phenolic levels. In addition, both our study and previous studies with north temperate and tropical species demonstrate that there is marked variation in algal phenolic levels within species from different locations. We conclude that high phenolic concentrations occur in species from both temperate and tropical regions, indicating that latitude alone is not a reasonable predictor of plant phenolic concentrations.     

Inborn errors of cobalamin metabolism: Effect of cobalamin supplementation in culture on methylmalonyl CoA mutase activity in normal and mutant human fibroblasts

We have examined the effect of addition of hydroxocobalamin to growth medium on the activity of the adenosylcobalamin-requiring enzyme methylmalonyl CoA mutase in normal human fibroblasts and in mutant human fibroblasts derived from patients with inherited methylmalonicacidemia. The mutant cell lines were assigned to four distinct genetic complementation groups (cbl A, cbl B, cbl C, and cbl D), each deficient in some step in the synthesis of adenosylcobalamin from hydroxocobalamin. After control cells were grown in cobalamin-supplemented medium, mutase holoenzyme activity increased markedly in a time- and concentration-dependent fashion. Growth in cobalamin-supplemented medium had no effect on mutase activity in some mutant lines belonging to the cbl B group, while activity increased severalfold in other cbl B mutants and in all cbl A, cbl C, and cbl D mutants examined, although mutase activity was still <10% of control. Comparison of mutase holoenzyme activity and total propionate pathway activity suggests that enhancement of mutase activity in mutant cells after cobalamin supplementation to values 5–10% of control may be sufficient to overcome the inherited metabolic block and to restore total pathway activity to normal.This work was supported in part by a research grant from the National Institutes of Health (AM 12579). H. F. W. is a recipient of a traineeship from the National Institutes of Health (T01-GM02299).     

Rhizosphere soil bacterial communities and nitrogen cycling affected by deciduous and evergreen tree species

Deciduous and evergreen trees differ in their responses to drought and nitrogen (N) demand. Whether or not these functional types affect the role of the bacterial community in the N cycle during drought remains uncertain. Two deciduous tree species (Alnus cremastogyne, an N₂‐fixing species, and Liquidambar formosana) and two evergreen trees (Cunninghamia lanceolata and Pinus massoniana) were used to assess factors in controlling rhizosphere soil bacterial community and N cycling functions. Photosynthetic rates and biomass production of plants, 16S rRNA sequencing and N‐cycling‐related genes of rhizosphere soil were measured. The relative abundance of the phyla Actinobacteria and Firmicutes was higher, and that of Proteobacteria, Acidobacteria, and Gemmatimondaetes was lower in rhizosphere soil of deciduous trees than that of evergreen. Beta‐diversity of bacterial community also significantly differed between the two types of trees. Deciduous trees showed significantly higher net photosynthetic rates and biomass production than evergreen species both at well water condition and short‐term drought. Root biomass was the most important factor in driving soil bacterial community and N‐cycling functions than total biomass and aboveground biomass. Furthermore, 44 bacteria genera with a decreasing response and 46 taxa showed an increased response along the root biomass gradient. Regarding N‐cycle‐related functional genes, copy numbers of ammonia‐oxidizing bacteria (AOB) and autotrophic ammonia‐oxidizing archaea (AOA), N₂ fixation gene (nifH), and denitrification genes (nirK, nirS) were significantly higher in the soil of deciduous trees than in that of the evergreen. Structural equation models explained 50.2%, 47.6%, 48.6%, 49.4%, and 37.3% of the variability in copy numbers of nifH, AOB, AOA, nirK, and nirS, respectively, and revealed that root biomass had significant positive effects on copy numbers of all N‐cycle functional genes. In conclusion, root biomass played key roles in affecting bacterial community structure and soil N cycling. Our findings have important implications for our understanding of plants control over bacterial community and N‐cycling function in artificial forest ecosystems.     

G4-quadruplex-binding proteins: review and insights into selectivity

There are over 700,000 putative G4-quadruplexes (G4Qs) in the human genome, found largely in promoter regions, telomeres, and other regions of high regulation. Growing evidence links their presence to functionality in various cellular processes, where cellular proteins interact with them, either stabilizing and/or anchoring upon them, or unwinding them to allow a process to proceed. Interest in understanding and manipulating the plethora of processes regulated by these G4Qs has spawned a new area of small-molecule binder development, with attempts to mimic and block the associated G4-binding protein (G4BP). Despite the growing interest and focus on these G4Qs, there is limited data (in particular, high-resolution structural information), on the nature of these G4Q-G4BP interactions and what makes a G4BP selective to certain G4Qs, if in fact they are at all. This review summarizes the current literature on G4BPs with regards to their interactions with G4Qs, providing groupings for binding mode, drawing conclusions around commonalities and highlighting information on specific interactions where available.