Hypomethylation of DNA during differentiation of friend erythroleukemia cells

DNA from mammalian cells has been shown to contain significant amounts of 5-methyl cytosine resulting from enzymatic transfer of methyl groups from s-adenosylmethionine to cytosine residues in the DNA polymer. The function of this modification is not known. We have found that DNA synthesized during chemically induced differentiation of friend erythroleukemia cells is hypomethylated, as measured by its ability to accept methyl groups transferred by homologous DNA methyltransferases in vitro. The extent of hypomethylation detected by this sensitive method is small, a decrease of less than 1.6 percent in 5-methylcytosine content. Hypomethylated DNA can be isolated from friend erythroleukemia cells grown in the presence of dimethyl sulfoxide, butyrate, hexamethylene-bis- acetamide, pentamethylene-bis acetamide, and ethionine. However, hypomethylated DNA is found only under conditions where differentiation is actually induced. DNA isolated from cells of a dimethyl sulfoxide- resistant subclone grown in the presence of that agent is not hypomethylated, although DNA of these cells becomes hypomethylated after growth in the presence of inducers that can trigger their differentiation. We also find that the DNA of friend erythroleukemia cells does not become hypomethylated when the cells are exposed to inducing agents in the presence of substances that inhibit differentiation. These results suggest a close link between genome modification by methylation and differentiation of friend erythroleukemia cells.     

Influence of Cobalt on Fermentative Methylation

Streptomyces rishiriensis produces at least five closely related antibiotics. Strain selection yielded a culture producing only the most active component, coumermycin A. Hydrolysis of this antibiotic by barium hydroxide yielded both 5-methyl-pyrrole-2-carboxylic acid and pyrrole-2-carboxylic acid, which could be separated by paper chromatography. Coumermycin A was thus shown to be two fractions, designated A(1) and A(2) depending upon the nature of the pyrrole carboxylic acid portion. The addition of cobalt to the fermentation medium at a level as low as 0.01 mug/ml shifted the fermentation exclusively to the production of coumermycin A(1). Other ions were ineffective, except nickel, whose activity could be explained by the presence of contaminating cobalt.     

Introducing systematics Agenda 2000

Systematics Agenda 2000 is an ambitious initiative emanating from biological systematists in the USA. It proposes an intensive international programme over a 25 year period;-(i) to discover, describe and inventory global species diversity, (ii) to synthesize the resulting data into a predictive classification, and (iii) to develop an appropriate information system to handle the resulting information.     

Prescribed burning in a Eucalyptus woodland suppresses fruiting of hypogeous fungi, an important food source for mammals

Fruit bodies of hypogeous fungi are an important food source for many small mammals and are consumed by larger mammals as well. A controversial hypothesis that prescribed burning increases fruiting of certain hypogeous fungi based on observations in Tasmania was tested in the Australian Capital Territory to determine if it applied in a quite different habitat. Ten pairs of plots, burnt and nonburnt, were established at each of two sites prescribe-burnt in May 1999. When sampled in early July, after autumn rains had initiated the fungal fruiting season, species richness and numbers of fruit bodies on the burnt plots were extremely low: most plots produced none at all. Both species richness and fruit body numbers were simultaneously high on nonburnt plots. One of the sites was resampled a year after the initial sampling. At that time species richness and fruit body abundance were still significantly less on burnt plots than on nonburnt, but a strong trend towards fungal recovery on the burnt plots was evident. This was particularly so when numbers of fruit bodies of one species, the hypogeous agaric Dermocybe globuliformis, were removed from the analysis. This species strongly dominated the nonburnt plots but was absent from burnt plots in both years. The trend towards recovery of fruit body abundance in the burnt plots one year after the burn was much more pronounced with exclusion of the Dermocybe data. The Tasmanian-based hypothesis was based mostly on the fruiting of two fire-adapted species in the Mesophelliaceae. Neither species occurred on our plots. Accordingly, the results and conclusions of the Tasmanian study cannot be extrapolated to other habitats without extensive additional study. Implications for management of habitat for fungi and the animals that rely on the fungi as a food source are discussed.     

Constrained (l-)-S-adenosyl-l-homocysteine (SAH) analogues as DNA methyltransferase inhibitors

Potent SAH analogues with constrained homocysteine units have been designed and synthesized as inhibitors of human DNMT enzymes. The five membered (2S,4S)-4-mercaptopyrrolidine-2-carboxylic acid, in 1a, was a good replacement for homocysteine, while the corresponding six-member counterpart was less active. Further optimization of 1a, changed the selectivity profile of these inhibitors. A Chloro substituent at the 2-position of 1a, compound 1d, retained potency against DNMT1, while N⁶ alkylation, compound 7a, conserved DNMT3b2 activity. The concomitant substitutions of 1a at both 2- and N⁶ positions reduced activity against both enzymes.     

Ecological relationships among hypogeous fungi and trees: inferences from association analysis integrated with habitat modeling

Association analyses by contingency tables and generalized linear modeling were compared to infer relationships among hypogeous (belowground-fruiting) ectomycorrhizal fungi and potential host tree species from 136 study plots in forested habitats in southeastern mainland Australia. Results from both types of statistical approaches were highly congruent. As with previous experimental studies, no exclusive fungus-host tree associations were identified. However, the likelihood of occurrence of some species of fungi increased significantly in the presence of particular host tree species, suggesting fungal host preference or shared habitat preferences. Similarly, while most associations among fungal species were nonsignificant, a few taxa were more likely to be found in the presence of certain others. These were termed positively associated and are thought to share common climatic and microhabitat requirements or host preferences. In contrast, other combinations of fungal species were negatively associated with one another, perhaps indicating different habitat preferences. Furthermore, the finding that some fungi occurred more frequently in the presence of certain tree species provides a starting point for selection of compatible host-fungus combinations that could be used for forest nursery and restoration applications.     

Microbial and chemical transformations of some 12,13-epoxytrichothec-9,10-enes.

Resting cells of Streptomyces griseus, Mucor mucedo, and a growing culture of Acinetobacter calcoaceticus when mixed with compounds related to 12,13-epoxytrichothec-9-ene-4beta,15-diacetoxy-3alpha-ol(anguidine) produced a series of derivatives that were either partially hydrolyzed or selectively acylated. These derivatives showed marked differences in activities as assayed by antifungal and tissue culture cytotoxicity tests.