Primary structure of the herpesvirus saimiri genome.

This report describes the complete nucleotide sequence of the genome of herpesvirus saimiri, the prototype of gammaherpesvirus subgroup 2 (rhadinoviruses). The unique low-G + C-content DNA region has 112,930 bp with an average base composition of 34.5% G + C and is flanked by about 35 noncoding high-G + C-content DNA repeats of 1,444 bp (70.8% G + C) in tandem orientation. We identified 76 major open reading frames and a set of seven U-RNA genes for a total of 83 potential genes. The genes are closely arranged, with only a few regions of sizable noncoding sequences. For 60 of the predicted proteins, homologous sequences are found in other herpesviruses. Genes conserved between herpesvirus saimiri and Epstein-Barr virus (gammaherpesvirus subgroup 1) show that their genomes are generally collinear, although conserved gene blocks are separated by unique genes that appear to determine the particular phenotype of these viruses. Several deduced protein sequences of herpesvirus saimiri without counterparts in most of the other sequenced herpesviruses exhibited significant homology with cellular proteins of known function. These include thymidylate synthase, dihydrofolate reductase, complement control proteins, the cell surface antigen CD59, cyclins, and G protein-coupled receptors. Searching for functional protein motifs revealed that the virus may encode a cytosine-specific methylase and a tyrosine-specific protein kinase. Several herpesvirus saimiri genes are potential candidates to cooperate with the gene for saimiri transformation-associated protein of subgroup A (STP-A) in T-lymphocyte growth stimulation.     

Effects of promoters on the enhancement of pectin methyl esterase expression inAspergillus niger

Summary A pectin methylesterase-encoding gene (pmeA)_has been cloned and transformed intoA. niger wild-type NRRL3. Transformants produced 20-fold more PME than the host strain. For studying the effects of different promoters on thepmeA expression two novel plasmids were constructed, in which thepmeA promoter was replaced by efficient promoters such as theA. nidulans glyceraldehyde-3-phosphate dehydrogenase (pK45) or theA. oryzae -amylase (pK61) promoter. The highest level of PME expression was achieved with theA. oryzae -amylase promoter, reaching a 200-fold increase compared to the production by the host strain.     

PHENOTYPIC PLASTICITY IN THE SAILFIN MOLLY,POECILIA LATIPINNA (PISCES: POECILIIDAE). II. LABORATORY EXPERIMENT

Field studies indicate that the influence of environmental factors on growth rate and size and age at maturity in sailfin mollies (Poecilia latipinna) is inconsistent over time and suggest that the marked interdemic variation in male body size in this species is the result of genetic variation. However, the role of specific environmental factors in generating phenotypic variation must be studied under controlled conditions unattainable in nature. We raised newborn sailfin mollies from four populations in laboratory aquaria under all possible combinations of two temperatures, three salinities, and two food levels to examine explicitly the influence of these environmental factors. Males were much less susceptible than females to temperature variation and were generally less plastic than females in terms of all three traits. Members of both sexes matured at larger sizes and at later ages in less saline and in cooler environments. Food levels were not sufficiently different to affect the traits we studied. The effects of temperature and salinity were not synergistic. Males from different populations exhibited different average ages and sizes at maturity, but females did not. The magnitudes of the effects we found were not substantial enough to account for the consistent interdemic differences in male and female body size that have been observed previously. Our results also indicate that no single environmental factor is solely responsible for the environmental effects observed in field experiments on growth and development. These studies, together with other work, indicate that the strongest sources of interdemic variation are genetic differences in males and differences in postmaturation growth and survivorship in females.     

A morphologic and immunofluorescent analysis of primary guinea pig glomerular cell types grown in chemically defined media. Evidence for clonal growth and cell differentiation

Primary kidney guinea pig glomerular cells were successfully grown in chemically defined media containing insulin, transferrin, and fibronectin or glycylhistidyllysine and fibronectin. Morphologic analysis of glomerular cells grown in either of these chemically defined media provided identical results with respect to cell growth properties and cell types involved. Electron microscopic studies of glomeruli early after they had been placed in culture showed definite evidence of "dedifferentiation" of some glomerular cells. Most glomerular cells in later cultures were undifferentiated. However, since electron microscopic analyses of glomeruli in confluent cultures demonstrated that the majority of cells in culture grow from the epithelial side of the glomerular basement membrane, we suggest that these cells were some form of epithelial cell. This conclusion was further strengthened by the fact that cells resembling well differentiated glomerular epithelial cells were seen in cultures of glomeruli grown in chemically defined media; these cells have never been observed in glomeruli grown in calf serum. Fluorescent microscopy of cell stained with the mitochondrial stain rhodamine 123 allowed identification of several glomerular cell types according to distribution, number, and morphology of mitochondria. Similarly, indirect immunofluorescent microscopy studies using antibodies to fibronectin or laminin provided evidence that glomerular cells separated into cell types according to mitochondrial staining properties were unique biochemically. Using these histochemical criteria it was possible to demonstrate that certain of the glomerular cell types could be selectively grown by addition of the enzyme galactose oxidase to the media. Analysis of our morphologic and histochemical results suggests the possibility that clonal growth and differentiation of glomerular epithelial cells occurs when glomeruli are placed in chemically defined media, and our results are compatible with the hypothesis that either "stem cells" or "dedifferentiated" cells are the primary cells dividing in culture.     

Treadmilling of actin at physiological salt concentrations: An analysis of the critical concentrations of actin filaments

Treadmilling of actin was investigated at physiological salt concentrations (100 mm-KCl, 0.5 to 2.0 mm-MgCl₂, 200 μm-ethyleneglycol-bis(β-aminoethyl ether)N,N′-tetraacetic acid or 50 μm-CaCl₂ at 37 °C. The concentration at which monomers bind to the lengthening end of filaments with the same rate as subunits are released (low critical concentration c₁ was determined by mixing unmodified actin filaments with various concentrations of monomeric actin labeled with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. Above a monomeric actin concentration of about 0.12 μm, incorporation of actin molecules into filaments was detected, whereas below this concentration no incorporation was found (c₁ = 0.12 μm). Combination of various concentrations of labeled monomers with labeled filaments permitted determination of the net critical concentration ($\text{c}{}_1$) at which filaments lengthen at one end with the same rate as they shorten at the other end ($\text{c}{}_1\text{= 0.16}\text{μm}$). A lower limit of the high critical concentration at the shortening end (c_h) was estimated by measuring the release of subunits from labeled filaments in the presence of various concentrations of unlabeled monomers (c_h >0.5 μm). The differences in the three critical concentrations demonstrate that under physiological conditions actin filaments lengthen at one end by, on the average, one subunit during the time that four association reactions take place at the two ends (efficiency parameters $\text{s =}\text{1}\text{4}$). The small difference between the low and the net critical concentration suggests that the rates of both association and dissociation are considerably greater at the lengthening end than at the shortening end of actin filaments.     

Identification of two nuclear polyhedrosis viruses from the cabbage moth, Mamestra brassicae (Lepidoptera: Noctuidae)

Two nuclear polyhedrosis viruses from the cabbage moth Mamestra brassicae found in two geographical areas in Europe have been characterized and compared. These two virus isolates have similar biological activities and have the same host range. The two M. brassicae nuclear polyhedrosis viruses can be distinguished by restriction endonuclease analysis of their DNA. They appear to be distinct but related virus strains.     

GD (-) Aachen,a new variant of deficient glucose-6-phosphate dehydrogenase

Summary A deficient G-6PD variant was discovered in 4 males of one family from north-western Germany. Five generations of this family could be studied.The deficient G-6PD was a new variant, called Gd (-) Aachen. Its main characteristics are the following: severe enzyme deficiency in erythrocytes (3% of normal), contrasting with an almost normal activity in leukocytes; normal molecular specific activity (i.e., normal ratio enzyme activity/cross-reacting material); slow mobility in starch gel electrophoresis (92–94% of normal); increased Michaelis constant for glucose-6-phosphate (60–70 M) and NADP⁺ (20–25 M); decreased inhibition constant by NADPH with respect to NADP⁺ (7 M); increased inhibition by ATP; normal utilization of the substrate analogues; slightly biphasic pH curve; thermal instability, and normal activation energy of the enzymatic reaction.The relationships between the hematologic disorders (severe and frequent hemolytic crises) and the unfavorable kinetic modifications are discussed.with the technical assistance of Joelle Marie and Dominique CottreauDedicated to Prof. Dr. H. Schonenberg, Aachen, on his 60th birthday. The first results of this work were presented in part at the Kongress der Deutschen Kinderärzte, München.