An inducible helix–Gly–Gly–helix motif in the N-terminal domain of histone H1e: A CD and NMR study

Knowledge of the structural properties of linker histones is important to the understanding of their role in higher-order chromatin structure and gene regulation. Here we study the conformational properties of the peptide Ac-EKTPVKKKARKAAGGAKRKTSG-NH(2) (NE-1) by circular dichroism and (1)H-NMR. This peptide corresponds to the positively charged region of the N-terminal domain, adjacent to the globular domain, of mouse histone H1e (residues 15-36). This is the most abundant H1 subtype in many kinds of mammalian somatic cells. NE-1 is mainly unstructured in aqueous solution, but in the presence of the secondary-structure stabilizer trifluoroethanol (TFE) it acquires an alpha-helical structure. In 90% TFE solution the alpha-helical population is approximately 40%. In these conditions, NE-1 is structured in two alpha-helices that comprise almost all the peptide, namely, from Thr17 to Ala27 and from Gly29 to Thr34. Both helical regions are highly amphipathic, with the basic residues on one face of the helix and the apolar ones on the other. The two helical elements are separated by a Gly-Gly motif. Gly-Gly motifs at equivalent positions are found in many vertebrate H1 subtypes. Structure calculations show that the Gly-Gly motif behaves as a flexible linker between the helical regions. The wide range of relative orientations of the helical axes allowed by the Gly-Gly motif may facilitate the tracking of the phosphate backbone by the helical elements or the simultaneous binding of two nonconsecutive DNA segments in chromatin.     

Morphological variation and cultural characteristics ofConiothyrium leucospermi associated with leaf spots of proteaceae

During an examination ofConiothyrium collections occurring on Proteaceae one species,C. leucospermi, was repeatedly encountered. However, it was not always possible to identify this species from host material alone, whereas cultural characteristics were found to be instrumental in its identification. Conidium wall ornamentation, which has earlier been accepted as crucial in species delimitation is shown to be variable on host material, making cultural comparisons essential. Using standard culture and incubation conditions,C. leucospermi is demonstrated to have a wide host range in the Proteaceae. In addition, microcyclic conidiation involving yeast-like budding from germinating conidia and hyphae in culture is newly reported for this species.     

Evaluation of bioassays to quantifyCylindrocaldium inocula in soil

Leaf discs provided better recovery ofCylindrocladium candelabrum from soil than stem, or twig segments. Leaf discs of eucalypt (Eucalyptus grandis), azalea (Rhododendron sp.), and geranium (Pelargonium sp.) were the best of seven plant baits evaluated. Twig segments of azalea and eucalypt also provided a high percentage of the pathogen recovery, whereas stem segments of pine seedlings (Pinus elliottii) proved unsatisfactory. Although slightly less effective, twig segments were easier to handle than leaf discs which were quickly decomposed in soil. Colonization of eucalypt twig segments byCylindrocladium spp. varied with inoculum level, soil moisture content, and incubation period. The highest percentage of recovery ofC. candelabrum (approx. 95%) was calculated at a field capacity moisture level of 155.9% after 75h of incubation.     

Xenocylindrocladium guianense andX. subverticillatum, two new species of hyphomycetes from plant debris in the tropics

Two new species of hyphomycetes,Xenocylindrocladium guianense andX. subverticillatum, are described from plant debris collected in French Guiana and Singapore, respectively. The genusXenocylindrocladium has thus far been known from one species,X. serpens, which was described from plant debris collected in Ecuador. The two new taxa are compared with and distinguished fromX. serpens based on morphology, cultural characteristics and phylogenetic analysis of DNA sequence data of the 5.8S rDNA with flanking ITS1 and ITS2 regions and the 5′ end of the β-tubulin gene. These species are also compared with other closely related hypocrealean taxa. Present collection data suggest that species ofXenocylindrocladium could be restricted to the tropics.     

The effects of long-term elevated UV-B on the growth and phenolics of field-grown silver birch (Betula pendula)

The effects of long‐term elevated UV‐B radiation on silver birch (Betula pendula Roth) seedlings were studied over three growing seasons in an outdoor experiment in Finland started 64 days after germination. One group of seedlings was exposed to a constant 50% increase in UV‐B_CIE radiation, which corresponds to 20–25% of ozone depletion; another group received a small increase in UV‐A radiation and a third (the control group) received ambient solar radiation. Changes in growth appeared during the third growing season; the stems of the UV‐B treated seedlings were thinner and their height tended to be shorter compared with that of the control seedlings. In contrast, there were no UV‐B effects on biomass, bud burst, bud dry weights, leaf area, rust frequency index or chlorophyll concentrations in any of the summers. During the three‐year study, the flavonols were significantly increased by the elevated UV‐B only in the first growing season. The responses varied greatly among individual compounds; the most induced were the quercetin glycosides, while the main flavonols, myricetins, were reduced by the UV‐A control treatment. In the second summer phenolic acids, such as 3,4′‐dihydroxypropiophenone‐3‐glucoside, neochlorogenic acid and 5‐coumarylquinic acid, were increased by the UV‐B treatment. In the third year, the constitutive concentrations of phenolics were not affected by the UV‐B treatment.     

Multi-disciplinary evidence of the Holocene history of a cultivated floodplain area in the wetlands of northern Colombia

An environmental reconstruction of the last 10,000 ¹⁴C years of a frequently flooded wetland ecosystem in the lower Magdalena valley in northern Colombia is presented, on the basis of a multi-disciplinary study of the sediments of the upper 15 m of the core from Boquillas (74°33'E, 9°7'N; 20 m a. s. l.). We used the following studies: pollen, lithology, organic structures, clay mineralogy, soil and sediment geochemistry, and δ¹³C values. The chronology is based on 13 AMS radiocarbon dates; the humic acid fractions were used in the case of seven samples. Pollen from local origin (swamps, open grass-rich vegetation, and gallery forest) show the development of the wetland area. River-transported pollen from a greater distance (dry forest, montane forest, Alnus) show changes in river activity and reflect large-scale changes of climatic conditions in the Momposina basin. From c. 10,010 to 9370 uncal B. P. (zone BQS-Ia) the river system was of high energy, as inferred by the lithological changes. The landscape was dominated by open grass-rich vegetation with gallery forest along the streams. A marked representation of Alnus and montane forest taxa indicate significant water transport and river dynamics. Climatic conditions were dry. From c. 9370-8430 uncal B. P. (zone BQS-Ib) wetlands were isolated from the main river system, and clayey sediments with kaolinite, smectite and illite as the main minerals accumulated in a lower-energy environment. Climatic conditions were dry and changes in the seasonal precipitation favoured the expansion of the gallery forest. From c. 8430 to 8040 uncal B. P. (zone BQS-Ic) low values of river-transported pollen indicate dry climatic conditions and open vegetation became more abundant. The flooding frequency of the Boquillas site diminished. From 8040 to 4900 uncal B. P. (zone BQS-Id) the Boquillas site was dominated by open vegetation with patches of gallery forest along the streams. Supply of river-transported allochthonous pollen (from many sources) was minimal. Clay minerals from the sediments suggest variable temperature and precipitation. From c. 4900 to 1550 uncal B. P. )zone BQS-II) the site was within the reach of the main river system as is the case today. Frequent flooding, coinciding with peaks of river-transported grains of Alnus and high sediment supply, point to high precipitation in the composite catchment area of the Magdalena, Cauca, San Jorge, and Cesár rivers. High values of phosphorous in the upper part of the core point to the presence of a pre-Hispanic civilization, approximately from 2000 uncal B. P. onward. Construction of an extensive drainage system allowed irrigation as well as drainage depending the annual cycle of precipitation. The landscape was significantly modified and allowed an extensive crop production on a system of raised fields. Received May 18, 2001 / Accepted June 15, 2001     

Effect of lateral far-red light supplementation on the growth and morphology of birch seedlings and its interaction with mineral nutrition

The effects of lateral far-red light (FR) and nutrient supply on the growth and nitrogen accumulation of silver birch (Betula pendula) seedlings were studied with the objective of testing the following hypotheses: (1) silver birch seedlings grow taller in response to increased FR irradiance received from the side; (2) this response is modulated by the nutritional status of the seedlings; and (3) an increase in lateral FR irradiance, and concomitant decrease in red to far-red photon ratio, affects the carbon and nitrogen economies of the seedlings. Two factorial experiments, each with two levels of mineral nutrient availability and two light treatments (background 'white' light with and without additional lateral FR simulating light reflected by neighbours in a sparse canopy) were done with small seedlings. The two experiments differed in PAR irradiance. The results of these experiments were that (1) stem elongation rate was increased by lateral FR addition, (2) there was no interaction between this effect and the nutritional status of the seedlings, and (3) neither whole plant accumulation of nitrogen nor dry weight increment was affected by lateral FR under either mineral nutrient supply regime.     

[NiFe] hydrogenase from Desulfovibrio desulfuricans ATCC 27774: gene sequencing, three-dimensional structure determination and refinement at 1.8 Å and modelling studies of its interaction with the tetrahaem cytochrome c 3

The primary and three-dimensional structures of a [NiFe] hydrogenase isolated from D. desulfitricans ATCC 27774 were determined, by nucleotide analysis and single-crystal X-ray crystallography. The three-dimensional structural model was refined to R=0.167 and Rfree=0.223 using data to 1.8 A resolution. Two unique structural features are observed: the [4Fe-4S] cluster nearest the [NiFe] centre has been modified [4Fe-3S-3O] by loss of one sulfur atom and inclusion of three oxygen atoms; a three-fold disorder was observed for Cys536 which binds to the nickel atom in the [NiFe] centre. Also, the bridging sulfur atom that caps the active site was found to have partial occupancy, thus corresponding to a partly activated enzyme. These structural features may have biological relevance. In particular, the two less-populated rotamers of Cys536 may be involved in the activation process of the enzyme, as well as in the catalytic cycle. Molecular modelling studies were carried out on the interaction between this [NiFe] hydrogenase and its physiological partner, the tetrahaem cytochrome c3 from the same organism. The lowest energy docking solutions were found to correspond to an interaction between the haem IV region in tetrahaem cytochrome c3 with the distal [4Fe-4S] cluster in [NiFe] hydrogenase. This interaction should correspond to efficient electron transfer and be physiologically relevant, given the proximity of the two redox centres and the fact that electron transfer decay coupling calculations show high coupling values and a short electron transfer pathway. On the other hand, other docking solutions have been found that, despite showing low electron transfer efficiency, may give clues on possible proton transfer mechanisms between the two molecules.     

Electric Field-Driven Disruption of a Native β-Sheet Protein Conformation and Generation of a Helix-Structure

We demonstrate that an external constant electric field is able to modify the secondary structure of a protein and induce a transition from a β-sheet into a helix-like conformation. This dramatic change is driven by a global rearrangement of the dipole moments at the amide planes. We also predict electric-field-induced modifications of the intermediate states of the protein.