Basis for slow growth on the non-fermentable substrates by a Saccharomyces cerevisiae mutant UV-sensitive for rho- production.

Summary The mutant uvs 72 of Saccharomyces cerevisiae UV-sensitive for rho^- production displays slower growth on media containing non-fermentable carbon sources such as glycerol or lactate. The slower growth on glycerol is not due to any deficiency in glycerol catabolism or mitochondrial oxidative phosphorylation. No modifications of the sensitivity to ethidium bromide of the mitochondrial ATPase activity could be detected. A mathematical model is presented which accounts for slower growth of uvs 72 on the sole basis of the continuous and elevated rho^- production in the mutant strain. This model, which estimates the rate of mutation from the rate of growth and vice versa, has been verified experimentally in the case of uvs 72. The model has been generalised, so that it can be used for any microbial population subject to constant and high rates of any type of mutation providing that the mutant is stable, and either unable to grow or able to grow at this own rate different from that of the parental strain.Abbreviations UV ultraviolet light (254 nm) - YG, YD, YL, YDG culture media containing respectively 3% glycerol, 1% glucose, 3% lactate and 0.1% glucose plus 3% glycerol This paper is number 1495 of the EURATOM Biology Division     

Callus and shoot formation in organ and tissue cultures of Hedera helix L., English ivy

When shoots of young plants of hemp (Cannabis sativa L.) and spinach (Spinacea oleracea L.) were cultured as cuttings and allowed to regenerate advenitious roots, ca. 80–85% became female (formed pistillate flowers) regardless of whether the leaves were left on the plants or were cut off (except for the 2–3 uppermost ones) after the beginning of adventitious-root formation. But when the leaves were cut off and the cuttings treated with gibberellic acid (GA₃, 25 mg/l) ca. 77–80% of the plants became male (formed staminate flowers). The result was quite similar when roots and leaves of young hemp plants were removed at the same time and the cuttings treated with GA₃. It is suggested that the leaves play an essential role in sex expression in hemp and spinach and that this role is related to gibberellin synthesis in the leaves.     

Electron spin resonance studies of an animal model of human congenital myotonia: Increased erythrocyte membrane fluidity in rats with 20,25-diazacholesterol-induced myotonia

Summary Electron spin resonance experiments have been performed on erythrocyte membranes from rats with myotonia induced by treatment with 20,25-diazacholesterol. The results suggest that erythrocyte membranes in this animal model of human congenital myotonia possess a highly significantly increased surface membrane fluidity compared to that of controls. Alterations in the physical state of membrane proteins were not apparent. These findings, also present in human congenital myotonia [Butterfield, Chesnut, Roses & Appel, 1976,Nature (London)263:159; Butterfield, 1977 (Submitted for publication)], strengthen the concepts that increased membrane fluidity is associated with the presence of myotonia and that congenital myotonia may be a diffuse membrane disease.     

EVOLUTIONARY HISTORY OF NORTHERN HEMISPHERE NUCELLA (GASTROPODA,MURICIDAE): MOLECULAR,MORPHOLOGICAL, ECOLOGICAL,AND PALEONTOLOGICAL EVIDENCE

By combining data from a variety of sources we explore patterns of evolution and speciation in Nucella, a widely studied genus of shallow-water marine neogastropods. We present a hypothesis of phylogenetic relationships for all of the currently recognized species of northern hemisphere Nucella, based on an analysis of 718 base pairs of nucleotide sequence from the mitochondrial cytochrome b gene. The order of appearance of species in the fossil record is congruent with this hypothesis. The topology of the inferred phylogeny of Nucella, coupled with ecological, morphological, and fossil evidence, was used to address three main questions: (1) At what time and by which route was the North Atlantic invaded from the North Pacific compared to prior studies of the trans-Arctic interchange? (2) Do patterns of molecular variation within species corroborate the importance of climatic cycles in driving speciation in north temperate marine animals? (3) Was radiation in the direction of increased or decreased ecological specialization, body size, or vulnerability to predation? Molecular evidence confirmed that the sole North Atlantic species, N. lapillus, arose from a North Pacific ancestor. Biogeographic and paleontological evidence supported the dispersal of Nucella, and perhaps other interchange species, via the Eurasian Arctic. Rather intriguingly, the linkage of N. lapillus to a western as opposed to eastern Pacific clade, and the biogeographic origins of the eastern Pacific species, parallel closely similar patterns observed in another genus of rocky-shore gastropods, Littorina. This congruence, in conjunction with information on the climatic and geographic histories of the region, as well as the geographic arrangement of mtDNA haplotypes within Nucella species, supports a model of speciation in Nucella driven by cycles of climatic amelioration and deterioration that began during the Miocene. Calibrations from the fossil record of Nucella suggest that third position transitions and transversions accrue at a rate of 3–4% and 0.5% respectively per million yr. This supports an early participation by Nucella in the trans-Arctic interchange, as suggested by paleobiogeographic studies. Consistent with the unstable taxonomic history of species of Nucella, we found few nonmolecular traits to be phylogenetically informative. Among North Pacific species, more recently derived species (N. canaliculata and the N. emarginata clade) were more ecologically specialized (narrower diet and habitat range). Consistent with extensive intraspecific variation, shell traits were quite labile evolutionarily: neither overall size nor development of antipredatory traits exhibited consistent evolutionary trends over the history of the genus. Nurse eggs (unfertilized eggs consumed by developing embryos) were an ancestral trait that was lost evolutionarily in the two clades that also exhibited increased body size, suggesting that these two life-history traits may be coupled. The reduced number of chromosomes in N. lapillus is clearly a derived state and is consistent with White's (1978) observations on chromosome evolution in other clades.     

INTRASPECIFIC MOLECULAR PHYLOGENY IN THE YELLOW WARBLER (DENDROICA PETECHIA), AND IMPLICATIONS FOR AVIAN BIOGEOGRAPHY IN THE WEST INDIES

A phylogenetic analysis of mitochondrial DNA (mtDNA) restriction sites was used to examine the evolutionary history of populations of yellow warbler (Dendroica petechia) sampled from North America, Central America, South America, and the West Indies. Thirty-seven haplotypes were identified, and only one was found in more than one of these regions. Estimated sequence divergence among haplotypes ranged from 0.14 to 3.17%, and mtDNAs from North American migratory populations clearly were differentiated from those of most tropical sedentary populations. Parsimony analysis of haplotypes suggested multiple colonizations of the West Indies archipelago and of individual Caribbean islands. The inference of multiple colonizations has important implications for studies of avian ecology and evolution in this region.     

Genetic Studies of Sulfadiazine-resistant and Methionine-requiring Neisseria Isolated From Clinical Material

Deoxyribonucleate (DNA) preparations were extracted from Neisseria meningitidis (four isolates from spinal fluid and blood) and N. gonorrhoeae strains, all of which were resistant to sulfadiazine upon primary isolation. These DNA preparations, together with others from in vitro mutants of N. meningitidis and N. perflava, were examined in transformation tests by using as recipient a drug-susceptible strain of N. meningitidis (Ne 15 Sul-s Met⁺) which was able to grow in a methionine-free defined medium. The sulfadiazine resistance typical of each donor was introduced into the uniform constitution of this recipient. Production of p-aminobenzoic acid was not significantly altered thereby. Transformants elicited by DNA from the N. meningitidis clinical isolates were resistant to at least 200 μg of sulfadiazine/ml, and did not show a requirement for methionine (Sul-r Met⁺). DNA from six strains of N. gonorrhoeae, which were isolated during the period of therapeutic use of sulfonamides, conveyed lower degrees of resistance and, invariably, a concurrent methionine requirement (Sul-r/Met⁻). The requirement of these transformants, and that of in vitro mutants selected on sulfadiazine-agar, was satisfied by methionine, but not by vitamin B₁₂, homocysteine, cystathionine, homoserine, or cysteine. Sul-r Met⁺ and Sul-r/Met⁻ loci could coexist in the same genome, but were segregated during transformation. On the other hand, the dual Sul-r/Met⁻ properties were not separated by recombination, but were eliminated together. DNA from various Sul-r/Met⁻ clones tested against recipients having nonidentical Sul-r/Met⁻ mutant sites yielded Sul-s Met⁺ transformants. The met locus involved is genetically complex, and will be a valuable tool for studies of genetic fine structure of members of Neisseria, and of genetic homology between species.     

Spermidine/spermine N(1)-acetyltransferase-1 binds to hypoxia-inducible factor-1alpha (HIF-1alpha) and RACK1 and promotes ubiquitination and degradation of HIF-1alpha

Hypoxia-inducible factor-1 (HIF-1) is a master regulator of oxygen homeostasis that controls the expression of genes encoding proteins that play key roles in angiogenesis, erythropoiesis, and glucose/energy metabolism. The stability of the HIF-1alpha subunit is regulated by ubiquitination and proteasomal degradation. In aerobic cells, O(2)-dependent prolyl hydroxylation of HIF-1alpha is required for binding of the von Hippel-Lindau tumor suppressor protein VHL, which then recruits the Elongin C ubiquitin-ligase complex. SSAT2 (spermidine/spermine N-acetyltransferase-2) binds to HIF-1alpha and promotes its ubiquitination/degradation by stabilizing the interaction of VHL and Elongin C. Treatment of cells with heat shock protein HSP90 inhibitors induces the degradation of HIF-1alpha even under hypoxic conditions. HSP90 competes with RACK1 for binding to HIF-1alpha, and HSP90 inhibition leads to increased binding of RACK1, which recruits the Elongin C ubiquitin-ligase complex to HIF-1alpha in an O(2)-independent manner. In this work, we demonstrate that SSAT1, which shares 46% amino acid identity with SSAT2, also binds to HIF-1alpha and promotes its ubiquitination/degradation. However, in contrast to SSAT2, SSAT1 acts by stabilizing the interaction of HIF-1alpha with RACK1. Thus, the paralogs SSAT1 and SSAT2 play complementary roles in promoting O(2)-independent and O(2)-dependent degradation of HIF-1alpha.