Induction of a cellular enzyme for energy metabolism by transforming domains of adenovirus E1a.

Brain creatine kinase is a major enzyme of cellular energy metabolism. It is overexpressed in a wide range of tumor cell lines and is used as a tumor marker. We reported recently that the promoter of the human gene has a strong sequence similarity to the adenovirus E2E promoter. This similarity suggested that the brain creatine kinase gene may be regulated by the viral activator E1a. Experiments reported here showed that both enzyme activity and mRNA levels were induced by the oncogenic products of the E1a region of adenovirus type 5, but unlike the viral E2E promoter, which is induced predominantly by E1a domain 3, brain creatine kinase induction required domains 1 and 2. These domains are important for transformation and for the association of E1a with the retinoblastoma gene product and other cellular proteins. The induction by an oncogene of a cellular gene for energy metabolism may be of significance for the metabolic events that take place after oncogenic activation.     

A structural model for the generation of continuous curvature on the surface of a retroviral capsid

The genome of a retrovirus is surrounded by a convex protein shell, or capsid, that helps facilitate infection. The major part of the capsid surface is formed by interlocking capsid protein (CA) hexamers. We report electron and X-ray crystallographic analysis of a variety of specimens assembled in vitro from Rous sarcoma virus (RSV) CA. These specimens all contain CA hexamers arranged in planar layers, modeling the authentic capsid surface. The specimens differ only in the number of layers incorporated and in the disposition of each layer with respect to its neighbor. The body of each hexamer, formed by the N-terminal domain of CA, is connected to neighboring hexamers through C-terminal domain dimerization. The resulting layer structure is very malleable due to inter-domain flexibility. A helix-capping hydrogen bond between the two domains of RSV CA creates a pivot point, which is central to controlling their relative movement. A similar mechanism for the governance of inter-domain motion was recently described for the human immunodeficiency virus type 1 (HIV-1) capsid, although there is negligible sequence identity between RSV and HIV-1 CA in the region of contact, and the amino acids involved in creating the pivot are not conserved. Our observations allow development of a physically realistic model for the way neighboring hexamers can tilt out of plane, deforming the hexamer layer and generating the continuously curved surfaces that are a feature of all retroviral capsids.     

Resource partitioning in rhinolophoid bats revisited

We assessed the ecomorphological structure of a guild of rhinolophoid bats in a Malaysian rainforest first described by Heller and von Helversen (1989). These authors found that the distribution of echolocation call frequencies used by 12 syntopic species was more even than expected from allometric relationships or in randomly generated communities, and that the observed minimal ratio was greater than expected by chance alone. In this study we were able to expand their guild to 15 species, but in doing so it became apparent that call frequencies might be less evenly distributed across the total frequency range than previously proposed. We replicated Heller and von Helversen’s (1989) analyses with the full 15-species complement but were unable to support their suggestion that rhinolophoid bats exhibit resource partitioning through differences in frequency bands. We adopted a multivariate approach and incorporated measures of body size and wing morphology into the analysis. We used phylogenetic autocorrelation to ensure that the species were statistically independentand principal component analysis to describe the morphological space occupied by the 15 species in the community and four additional species representing the extremes of phenotypic variation. We derived interspecific Euclidean distances and tested the mean values and SDs of these distances against those of 100 guilds of ”synthetic” species created randomly within the principal component space. The guild of Rhinolophoidea was not distributed randomly in multivariate space. Instead we found evidence of morphological overdispersion of the most similar species, which suggests niche differentiation in response to competition. Less similar species were nearer in morphological space than expected, and we suggest this is a consequence of ecological constraints on parameter combinations. Despite this underdispersion, many of the more distant neighbours were evenly rather than randomly spaced or clumped in morphospace, suggesting that, given the environmental constraints on morphology, species in this guild do experience limits to their similarity. Finally, we tested the influence of the relative abundance of species on morphological displacement, and found no evidence that abundant, spatially correlated species reduce interspecific overlap in morphological space. Received: 10 April 1999 / Accepted: 28 February 2000     

Linking genotype to phenotype in a changing ocean: inferring the genomic architecture of a blue mussel stress response with genome‐wide association

A key component to understanding the evolutionary response to a changing climate is linking underlying genetic variation to phenotypic variation in stress response. Here, we use a genome‐wide association approach (GWAS) to understand the genetic architecture of calcification rates under simulated climate stress. We take advantage of the genomic gradient across the blue mussel hybrid zone (Mytilus edulis and Mytilus trossulus) in the Gulf of Maine (GOM) to link genetic variation with variance in calcification rates in response to simulated climate change. Falling calcium carbonate saturation states are predicted to negatively impact many marine organisms that build calcium carbonate shells – like blue mussels. We sampled wild mussels and measured net calcification phenotypes after exposing mussels to a ‘climate change’ common garden, where we raised temperature by 3°C, decreased pH by 0.2 units and limited food supply by filtering out planktonic particles >5 μm, compared to ambient GOM conditions in the summer. This climate change exposure greatly increased phenotypic variation in net calcification rates compared to ambient conditions. We then used regression models to link the phenotypic variation with over 170 000 single nucleotide polymorphism loci (SNPs) generated by genotype by sequencing to identify genomic locations associated with calcification phenotype, and estimate heritability and architecture of the trait. We identified at least one of potentially 2–10 genomic regions responsible for 30% of the phenotypic variation in calcification rates that are potential targets of natural selection by climate change. Our simulations suggest a power of 13.7% with our study's average effective sample size of 118 individuals and rare alleles, but a power of >90% when effective sample size is 900.     

Anaerobic metabolism of 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ) by human fecal flora

Incubation of the heterocyclic cooked food mutagen IQ with mixed human fecal microflora under anaerobic conditions yielded 2-amino-3,6-dihydro-3-methyl-7H-imidazo[4,5-f]quinoline-7-one (2) as the major detectable metabolite.     

Extraction, purification, identification and metabolism of 3'',5''-cyclic UMP, 3'',5''-cyclic IMP and 3'',5''-cyclic dTMP from rat tissues.

The large-scale extraction and partial purification of endogenous 3',5'-cyclic UMP, 3',5'-cyclic IMP and 3',5'-cyclic dTMP are described. Rat liver, kidney, heart, spleen and lung tissues were subjected to a sequential purification procedure involving freeze-clamping, perchlorate extraction, alumina and Sephadex ion-exchange chromatography and preparative electrophoresis. The samples thus obtained co-chromatographed with authentic cyclic UMP, cyclic IMP and cyclic dTMP on t.l.c. and h.p.l.c. and the u.v. spectra of the extracted samples were identical with those of the standards. Fast atom bombardment of the three cyclic nucleotide standards yielded mass spectra containing a molecular protonated ion in each case; mass-analysed ion kinetic-energy spectrometry ('m.i.k.e.s') of these ions produced a spectrum unique to the parent cyclic nucleotide. The extracted putative cyclic UMP, cyclic IMP and cyclic dTMP each produced a m.i.k.e.s. identical with that obtained with the corresponding cyclic nucleotide standard. Rat liver, heart, kidney, brain, intestine, spleen, testis and lung protein preparations were each found capable of the synthesis of cyclic UMP, cyclic IMP and cyclic dTMP from the corresponding nucleoside triphosphate, of the hydrolysis of these cyclic nucleotides and of their binding, with the exception that cyclic dTMP was not synthesized by the kidney preparation.     

Use of immobilized cells of the yeast,Saccharomycopsis lipolytica,for the production of citric acid

Biotechnology Letters - A strain ofSaccharomycopsis lipolytica was immobilized in polyacrylamide gel, and its ability to produce citric acid from glucose was examined. The immobilization procedure...     

Studies on the growth of the fetal sheep. The effects of reduction of placental size on hormone concentration in fetal plasma

Intrauterine growth retardation was induced in sheep by removal of endometrial caruncles before pregnancy. At a second operation catheters were implanted into the ewe and fetus at 105-135 days of pregnancy. Three groups of fetuses: low birthweight-for-dates (small caruncle) normal birthweight-for dates (normal sized caruncle) and controls have been compared. The concentration of ACTH (60 +/- 6.9 pg/ml) in the normal-sized caruncle fetuses were lower in the controls (144 +/- 4.7 pg/ml) or small caruncle fetuses (142 +/- 53 pg/ml). Basal cortisol concentrations were similar in the controls (7.3 +/- 1.2 ng/ml) and normal-sized caruncle fetuses (6.5 +/- 0.5 ng/ml) but those in the small caruncle fetuses were significantly higher (12.7 +/- 1.0 ng/ml, P less than 0.001). The concentration of insulin correlated with plasma glucose and the mean concentrations were 19.2 +/- 1.6 mu units/ml (controls), 8.4 +/- 2.6 mu units/ml (normal-sized caruncle) and 3.9 +/- 1.6 mu units/ml (controls), 8.4 +/- 2.6 mu units/ml (normal-sized caruncle) and 3.9 +/- 1.6 mu units/ml (small caruncle). Prolactin was significantly lower in the small caruncle fetuses (2.1 +/- 0.3 ng/ml) compared to the controls (66.6 19.4 ng/ml) or normal-sized caruncles (76.1 +/- 38 ng/ml) but growth hormone concentrations in the small caruncle.