Uracil-DNA glycosylase activities in hyperthermophilic micro-organisms

Abstract Hyperthermophiles exist in conditions which present an increased threat to the informational integrity of their DNA, particularly by hydrolytic damage. As in mesophilic organisms, specific activities must exist to restore and protect this template function of DNA. In this study we have demonstrated the presence of thermally stable uracil-DNA glycosylase activities in seven hyperthermophiles; one bacterial: Thermotoga maritima , and six archaeal: Sulfolobus solfataricus, Sulfolobus shibatae, Sulfolobus acidocaldarius, Thermococcus litoralis, Pyrococcus furiosus and Pyrobaculum islandicum . Uracil-DNA glycosylase inhibitor protein of the Bacillus subtilis bacteriophage PBS1 shows activity against all of these, suggesting a highly conserved tertiary structure between hyperthermophilic and mesophilic uracil-DNA glycosylases.     

Mutational spectrum in the neurofibromatosis type 2 gene in sporadic and familial schwannomas

Using a heteroduplex approach and direct sequencing, we have completed the screening of approximately 88% of the neurofibromatosis type 2 (NF2)-coding sequence of DNA extracted from 33 schwannomas from NF2 patients and from 29 patients with sporadic schwannomas. The extensive screening has resulted in the identification of 33 unique mutations. Similarly to other human genes, we have shown that the CpG sites are more highly mutable in the NF2 gene. The frequency, distribution, and types of mutations were shown to differ between the sporadic and familial tumors. The majority of the mutations resulted in protein truncation and were consistent with more severe phenotype, however three missense mutations were identified during this study and were all associated with milder manifestations of the disease. Received: 25 September 1995 / Revised: 19 December 1995     

Limitations to the inference of gene flow at regional geographic scales–an example from the Pieris napi group (Lepidoptera: Pieridae) in Europe

We used hierarchical and pairwise F-statistics to describe genetic differentiation and infer gene flow (M) on local and regional scales within and among parapatric European butterfly taxa in the Pieris napi (L.) group. Within-population allozyme variability is consistently high, and local effective population sizes are inferred to be in the thousands of individuals. The pairwise analysis yields an average neighbourhood area of radius 3.5 km. Among populations within most regions, differentiation is low and M > 2 effective individuals population^-1generation^-1. Pairwise comparisons within the brilannica group show a disjunction indicating that it is out of equilibrium, perhaps as a result of secondary contact between highland and lowland groups. Comparison between meridionalis groups on mainland Italy and Corsica yields M > 12; this is surely loo high and lack of equilibrium resulting from initial colonization is suspected. The hierarchical analysis indicates that 23 ≤0020M≤ 88 among the taxa napi, bryoniae and meridionalis that meet in hybrid zones; no effective gene flow barrier exists among them. This high estimate could also result from recent primary contact, but such a genetic barrier should produce the ‘edge effects' seen in population genetic simulations, and no evidence of this was found among geographically close samples of napi and bryoniae populations from Switzerland. Studies of gene flow among geographic regions are greatly limited by the equilibrium assumption, though studies of local differentiation are much less so. Population studies of gene flow on local scales at regional boundaries provide limited means of testing the equilibrium assumption, and both regional and local analyses provide testable predictions about local population structure. When the equilibrium assumption is not upheld, local patterns at regional boundaries can provide historical information about primary vs. secondary contact.     

Middle Cambrian pterobranchs and the Question: What is a graptolite?

The presence of distinct fusellar structure is taken as evidence to include a number of fossils from the Middle Cambrian to the Lower Ordovician of North America and Europe with the Pterobranchia. The dome of the pterobranchs and the prosicula of the planktic graptolites are contrasted and evidence is given for the re‐assignment of a number of well known dendroid graptolites to the pterobranchs. A non‐destructive method is described to reveal fusellar development of delicate hemichordate exoskeletons from shales. Rhabdotubus robustus n. sp. from the Czech Republic and ? Cephalodiscus sp. from the Wheeler Shale of North America are described as new Middle Cambrian pterobranchs.     

Soil Consumption of Two Groups of Semi-free-ranging Lemurs {Lemur catta and Lemur fulvus)

Soil consumption of two Malagasy lemurs, Lemur catta and Lemur fulvus was observed in an enclosure simulating a natural habitat at Duke University Primate Center (Durham, USA). Soil eaten by L. catta contained more Na than random soil samples. Concentrations of the four major minerals in soil eaten by L. fulvus did not differ from random samples. L. fulvus stopped soil ingestion in summer when they ate large amounts of foliage rich in K and Mg, also in winters after being fed with MgCl₂. It is suggested that, in this study, soil was consumed in relation to its mineral content and not for physical properties related to its structure. Some non physiological factors that might influence food selection are discussed.     

ATP binding and hydrolysis are essential to the function of the Hsp90 molecular chaperone in vivo.

Hsp90 is an abundant molecular chaperone essential to the establishment of many cellular regulation and signal transduction systems, but remains one of the least well described chaperones. The biochemical mechanism of protein folding by Hsp90 is poorly understood, and the direct involvement of ATP has been particularly contentious. Here we demonstrate in vitro an inherent ATPase activity in both yeast Hsp90 and the Escherichia coli homologue HtpG, which is sensitive to inhibition by the Hsp90-specific antibiotic geldanamycin. Mutations of residues implicated in ATP binding and hydrolysis by structural studies abolish this ATPase activity in vitro and disrupt Hsp90 function in vivo. These results show that Hsp90 is directly ATP dependent in vivo, and suggest an ATP-coupled chaperone cycle for Hsp90-mediated protein folding.     

cAMP and PMA enhance the effects of IGF-I in the proliferation of endometrial adenocarcinoma cell line HEC-1-A by acting at the G1 phase of the cell cycle

The present study was undertaken to determine whether endometrial cancer cell line HEC-1-A differ from nontransformed cells, in that the cAMP and protein kinase C pathways may enhance IGF-I effects in mitogenesis by acting at the G₁ phase of the cell cycle instead of G₀. Immunofluorescence staining of HEC-1-A cells using the proliferating cell nuclear antigen (PCNA) monoclonal antibody and flow cytometric analysis determined that HEC-1-A cells do not enter the G₀ phase of the cell cycle when incubated in a serum-free medium. Approximately 51% of the cells were in G₁, 12% were in S and 37% in G₂ phase of the cell cycle prior to treatment. Forskolin and phorbol-12-myristate 13-acetate (PMA) were used to stimulate cAMP production and protein kinase C activity, respectively. IGF-I, forskolin and PMA each increased (P <0.01) [³H]-thymidine incorporation in a dose and time dependent manner. The interaction of forskolin and PMA with IGF-I was then determined. Cells preincubated with forskolin or PMA followed by incubation with IFG-I incorporated significantly more (P <0.01) [³H]-thymidine into DNA than controls or any treatment alone. It is concluded that forskolin and, to a lesser extent, PMA exert their effect at the G₁ phase of the cycle to enhance IGF-I effects in cell proliferation.     

Regional variation in drivers of connectivity for two frog species (Rana pretiosa and R. luteiventris) from the U.S. Pacific Northwest

Comparative landscape genetics has uncovered high levels of variability in which landscape factors affect connectivity among species and regions. However, the relative importance of species traits versus environmental variation for predicting landscape patterns of connectivity is unresolved. We provide evidence from a landscape genetics study of two sister taxa of frogs, the Oregon spotted frog (Rana pretiosa) and the Columbia spotted frog (Rana luteiventris) in Oregon and Idaho, USA. Rana pretiosa is relatively more dependent on moisture for dispersal than R. luteiventris, so if species traits influence connectivity, we predicted that connectivity among R. pretiosa populations would be more positively associated with moisture than R. luteiventris. However, if environmental differences are important drivers of gene flow, we predicted that connectivity would be more positively related to moisture in arid regions. We tested these predictions using eight microsatellite loci and gravity models in two R. pretiosa regions and four R. luteiventris regions (n = 1,168 frogs). In R. pretiosa, but not R. luteiventris, connectivity was positively related to mean annual precipitation, supporting our first prediction. In contrast, connectivity was not more positively related to moisture in more arid regions. Various temperature metrics were important predictors for both species and in all regions, but the directionality of their effects varied. Therefore, the pattern of variation in drivers of connectivity was consistent with predictions based on species traits rather than on environmental variation.     

Population processes under the influence of disasters occurring independently of population size

Markov branching processes and in particular birth-and-death processes are considered under the influence of disasters that arrive independently of the present population size. For these processes we derive an integral equation involving a shifted and rescaled argument. The main emphasis, however, is on the (random) probability of extinction. Its distribution density satisfies an equation which can be solved numerically at least up to a multiplicative constant. In an example it is also found by simulation.