Cystatin domains in alpha-2-HS-glycoprotein and fetuin

We have found that chain A of alpha-2-HS-glycoprotein contains two cystatin domains that show closest similarity to those of kininogen. Most likely, the two proteins diverged after the primary duplication of a single cystatin domain as the two cystatin domains of alpha-2-HS-glycoprotein are more similar, especially in disulfide bonding, to the corresponding domains of kininogen than to each other. We also propose that the carboxyl-terminal (non-cystatin) parts of kininogen and alpha-2-HS-glycoprotein contain homologous segments. We suggest that alpha-2-HS-glycoprotein may act as an inhibitor of the cysteine proteinases responsible for bone resorption. We have also found that fetuin is closely related to alpha-2-HS-glycoprotein.     

Expression of polyoma virus middle-T antigen in Saccharomyces cerevisiae

The polyoma middle-T gene, lacking its intron, was inserted into a yeast expression plasmid containing the phosphoglycerate kinase promoter. Such plasmids transformed yeast at low frequency and these transformants expressed middle-T antigen at a level of approximately 0.1% cell protein. Furthermore, expression of this protein was frequently lost during growth in liquid culture and this loss of middle-T was accompanied by a twofold increase in the rate of growth. The spontaneous production of a truncated middle-T antigen, lacking the C terminus, was also observed; the expression of this protein did not inhibit the growth rate of the cells. Recovery and analysis of the expression plasmids encoding the truncated molecule showed that a single C X G base pair had been deleted from a run of nine consecutive C X G base pairs (Pyr nucleotide 1239--1247) within the middle-T coding region. This frame-shift mutation results in premature termination of the protein and loss of the strongly hydrophobic region of the molecule believed to be responsible for the membrane association of middle-T antigen.     

Electron microscopic studies of condensed mitotic chromosomes in the fission yeast Schizosaccharomyces pombe

By blocking cells in mitosis with the anti-fungal drug thiabendazole, it has been possible to carry out ultrastructural studies on the condensed chromosomes of the fission yeast, Schizosaccharomyces pombe. It is estimated that the DNA in these chromosomes is compacted approximately 1000-fold, and that the nucleoprotein density is similar to that of higher eukaryotic metaphase chromosomes. A basic structural component of the condensed chromosomes appears to be a 50-60 nm fibre, which is often visible in a loop configuration on the periphery of the chromatids. This is reminiscent of the 50-60 nm fibre loops which are frequently seen in preparations of metaphase chromosomes, and suggests that mechanisms of nucleoprotein folding may be similar in both lower and higher eukaryotes.     

A new species of Rhynchoidomonas Patton, 1910 (Kinetoplastida: Trypanosomatina) from Operophtera brumata (Lepidoptera: Geometridae)

Summary A new species of Rhynchoidomonas Patton was observed in a single adult male winter moth, Operophtera brumata (L.) from England. Intracellular amastigotes, and extracellular epimastigotes and trypomastigotes with an undulating membrane and free flagellum, were present. All stages had a large, reniform kinetoplast. As transmission of the flagellate between generations of winter moths by ingestion of infected faeces is a virtual impossibility, it is suggested that the flagellate's true host may have been a dipteran parasitoid and that an egg, surface-contaminated with the flagellate, was oviposited into or ingested by a winter moth larva. If the parasitoid had died, this flagellate infection could have been carried over to the adult moth. ac]19830601     

Selection for increased abdominal bristle number in Drosophila melanogaster with concurrent irradiation

Summary Replicate lines, each, with one hundred pairs of parents selected at 50% intensity, were derived from an inbred line. For twenty generations three lines were selected without irradiation and five with 1000 r X-rays per generation given to both females and males. After adjustment for level of crowding in the cultures, the final mean was 1.3 bristles higher in females and 1.0 bristles higher in males in the irradiated lines than in the unirradiated lines. In terms of phenotypic standard deviations in each sex in the base population, these total responses were 0.74 and 0.59 respectively. Radiation can induce mutations useful in increasing responses in selection programmes, but the average response attributable to radiation is small, and heterogeneity between replicate lines is to be expected.This work was carried out while B. H. was on leave from the New South Wales Department of Agriculture, and held a Commonwealth Research Studentship. Our thanks to Robin Hall and Tricia Brown for technical assistance, to Dr. B. L. Sheldon, C.S.I.R.O. Division of Animal Genetics, for providing the inbred line, and to the Radio-Therapy Unit, Royal Prince Alfred Hospital, for the use of their equipment.     

Effects of Population Size and Selection Intensity on Responses to Disruptive Selection in DROSOPHILA MELANOGASTER

Disruptive selection for sternopleural bristle number with opportunity for random mating was done in the four treatment combinations of two population sizes (40 pairs and 8 pairs of selected parents) and two selection intensities (1 in 40 and 1 in 2). In each generation, matings among selected parents were observed in a mating chamber, and progeny collected separately from each female parent. In the high number, high selection intensity treatment, divergence between the high and low parts ceased about generation 11. The isolation index increased rapidly to generation 3, but then fluctuated to termination of the population at generation 17. The overall isolation index was significant, indicating a real tendency to assortative mating. The failure of the isolation index to increase after generation 3 was attributed to lower average mating fitness of high males (due to inbreeding) and reduced receptivity of low females (due to a homozygous lethal gene with a large effect on sternopleural bristle number in heterozygotes). In the two low number treatments, isolation indices fluctuated from generation to generation with no obvious trends, and none of the overall isolation indices were significantly different from zero. The high number, low selection intensity treatment showed very little divergence, and one of the replicates showed, in contrast with expectation and the high number, high selection intensity treatment, a significant tendency to disassortative mating. Intense disruptive selection may lead to assortative mating.     

Metabolism of beta-methylaspartate by a pseudomonad

A bacterium was isolated from soil which utilizes threo-beta-methyl-l-aspartate, certain other amino acids, and a variety of organic substances as single energy sources. It is, or closely resembles, Pseudomonas putida biotype B. The ability of this organism to rapidly decompose such amino acids is dependent on inducible enzyme systems. Dialyzed cell-free extracts of this bacterium metabolize beta-methylaspartate only when catalytic amounts of alpha-ketoglutarate, or pyruvate, and pyridoxal phosphate are also present. The main products formed from beta-methylaspartate under these conditions are alpha-aminobutyrate, carbon dioxide, and alpha-ketobutyrate. When l-aspartate is substituted for beta-methylaspartate in this system, it is converted mainly to alanine and carbon dioxide. beta-Methyloxalacetate is decarboxylated, and the resulting alpha-ketobutyrate is converted enzymatically in the presence of glutamate to alpha-aminobutyrate which accumulates. The added keto acids are converted, in part, to the corresponding amino acids probably by transamination. The data indicate that beta-methylaspartate is converted to alpha-aminobutyrate, and aspartate to alanine, by a circuitous transamination-beta-decarboxylation-transamination sequence rather than by a direct beta-decarboxylation.